Oligonucleotide Composition and Method Thereof

Abstract

In particular, the present disclosure provides oligonucleotides, compositions, and methods useful for adenosine modification. In some embodiments, the present disclosure provides methods for treating various conditions, disorders, or diseases that may benefit from adenosine modification.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 341,407, filed May 12, 2022, which is incorporated by reference in its entirety. [Background technology]

[0002] background Oligonucleotides are useful in a variety of applications, such as therapeutic, diagnostic, and / or research applications. For example, oligonucleotides targeting various genes may be useful in treating conditions, disorders, or diseases associated with such target genes.

[0003] overview When used for various applications, oligonucleotide is typically prepared to have a base sequence that is complementary to a single stretch of consecutive nucleic acid bases in target nucleic acid.For example, in the case of target adenosine editing by ADAR, the base sequence of oligonucleotide is typically complementary to the base sequence of the part of target nucleic acid that contains target adenosine. Summary of the Invention [Means for solving the problem]

[0004] Surprisingly, the present invention demonstrates, inter alia, that an oligonucleotide has two or more domains, each domain independently has a base sequence complementary to a portion of target nucleic acid, and the two or more portions of target nucleic acid are independently separated by gaps.As demonstrated herein, such oligonucleotides surprisingly can provide various biological activities and are useful for various applications, such as target adenosine editing by ADAR.In some embodiments, the present disclosure provides an oligonucleotide comprising: A first domain; and Second Domain Including, a base sequence of the first domain is complementary to a first portion of a base sequence of a target nucleic acid; the base sequence of the second domain is complementary to a second portion of the base sequence of the target nucleic acid; An oligonucleotide is provided in which a first portion and a second portion of the base sequence of a target nucleic acid are separated by a gap.

[0005] The provided techniques, e.g., oligonucleotides, compositions, methods, etc., can provide various advantages. For example, in some embodiments, the provided techniques provide significantly expanded options for the base sequence of the oligonucleotide. For example, in the case of targeted adenosine editing, the base sequence of the oligonucleotide does not need to be complementary to a portion of the target nucleic acid within a certain range from the target adenosine (e.g., about 25 to 50 or more consecutive nucleosides including the target adenosine). A portion of the oligonucleotide, e.g., a first portion, may have a base sequence complementary to a distal portion of the target nucleic acid from the target adenosine. Alternatively or in addition, a shorter portion around the target adenosine may be targeted. In some embodiments, such a shorter portion avoids the destruction of one or more functional elements, e.g., motifs important for splicing, regulation, etc., close to the target adenosine (e.g., within about 5, 10, 15, 20, 30, 40, or 50 nucleosides from the target adenosine). Alternatively or additionally, the provided technology can improve various properties and / or activities, such as stability, cellular uptake, non-specific interactions, editing efficiency, etc., by providing the option of utilizing sequences complementary to portions away from the target adenosine. Alternatively or additionally, the provided technology allows or improves the editing of the target adenosine in complex structures, such as complex secondary RNA structures. Alternatively or additionally, the provided technology can provide different on / off rates. In some embodiments, the base sequence of the reference oligonucleotide for comparison is or includes a sequence complementary to the base sequence of only one portion of the target nucleic acid, the portion including the target adenosine, and is about 25-100, 30-80, 30-70, 30-60, 30-50, 25-50, or 25-40 nucleobases in length.

[0006] In some embodiments, the disclosure provides designed oligonucleotides and compositions thereof, where the oligonucleotides include modifications (e.g., modifications to nucleobase sugars and / or internucleotide linkages and patterns thereof) as described herein. In some embodiments, the techniques (compounds (e.g., oligonucleotides), compositions, methods, etc.) of the disclosure (e.g., oligonucleotides, oligonucleotide compositions, methods, etc.) are particularly useful for editing nucleic acids (e.g., site-specific editing in nucleic acids (e.g., editing of targeted adenosines)). In some embodiments, the techniques provided can significantly improve the efficiency of nucleic acid editing (e.g., modifying one or more A residues, such as converting A to I). In some embodiments, the disclosure provides techniques for editing in RNA (e.g., modifying A residues, e.g., converting A to I). In some embodiments, the disclosure provides techniques for editing in transcripts (e.g., mRNA) (e.g., modifying A residues, e.g., converting A to I). In particular, the provided technology offers the benefits of utilizing endogenous proteins, such as ADAR (adenosine deaminase acting on RNA) proteins (e.g., ADAR1 and / or ADAR2) to edit nucleic acids (e.g., to modify A (e.g., as a result of a G to A mutation)). One of skill in the art will appreciate that such utilization of endogenous proteins may avoid some challenges and / or provide various advantages compared to those technologies that require delivery of foreign components (e.g., proteins (e.g., engineered to bind to an oligonucleotide (and / or its duplex with a target nucleic acid) that results in a desired activity), nucleic acids encoding proteins, viruses, etc.).

[0007] In particular, in some embodiments, the oligonucleotides of the provided technology include useful sugar modifications and / or patterns thereof (e.g., the presence and / or absence of certain modifications), nucleobase modifications and / or patterns thereof (e.g., the presence and / or absence of certain modifications), internucleotide linkage modifications and / or stereochemistry and / or patterns thereof (e.g., chiral linkage phosphorus type, modification, and / or configuration (Rp or Sp), etc.), and when combined with one or more other structural elements (e.g., additional chemical moieties) described herein, may provide enhanced activity and / or various desirable properties (e.g., enhanced efficiency of nucleic acid editing, enhanced selectivity, enhanced stability, enhanced cellular uptake, reduced immune stimulation, reduced toxicity, improved distribution, improved affinity, etc.). In some embodiments, the provided oligonucleotides provide enhanced stability, for example, as compared to oligonucleotides having a higher percentage of natural RNA sugars utilized for adenosine editing. In some embodiments, the provided oligonucleotides provide enhanced activity (e.g., adenosine editing activity). In some embodiments, provided oligonucleotides provide high selectivity, e.g., in some embodiments, provided oligonucleotides provide for selective modification of a target adenosine in a target nucleic acid over other adenosines in the same target nucleic acid (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20-fold or more modification of the target adenosine relative to another adenosine, or all other adenosines in the target nucleic acid).

[0008] In particular, the disclosure provides designed oligonucleotides and compositions that have improved properties and / or activity compared to reference oligonucleotides and compositions (e.g., those described herein or reported in the art). For example, in some embodiments, the provided oligonucleotides and compositions may provide improved stability, pharmacokinetic properties, pharmacodynamic properties, and / or improved activity (e.g., with respect to A to I editing). Various designed oligonucleotides and compositions are described herein. For example, in some embodiments, the disclosure provides oligonucleotides and compositions thereof (e.g., chiral controlled oligonucleotide compositions thereof) that include sugar modifications at the 5' and 3' ends (e.g., 2'-OR modifications, where R is an optionally substituted C 1~6In some embodiments, the first several (e.g., 1, 2, 3, 4, or more; in some embodiments, 3 or more) nucleosides independently comprise a sugar modification, such as a cyclic sugar (e.g., an alkyl sugar) (e.g., 2'-OMe, 2'-MOE, etc.), a bicyclic sugar (e.g., an LNA sugar, a cEt sugar, etc.). In some embodiments, the first several (e.g., 1, 2, 3, 4, or more; in some embodiments, 3 or more) nucleosides and / or the last several (e.g., 1, 2, 3, 4, or more; in some embodiments, 3 or more) nucleosides independently comprise a sugar modification. In some embodiments, the first three or more and the last three or more nucleosides independently comprise a sugar modification. In some embodiments, one or more internucleotide linkages attached to such nucleosides are non-negatively charged internucleotide linkages (e.g., a phosphorylguanidine internucleotide linkage such as n001). In some embodiments, both the first and last internucleotide linkages are independently non-negatively charged internucleotide linkages. In some embodiments, both the first and last internucleotide linkages are independently phosphorylguanidine internucleotide linkages. In some embodiments, both the first and last internucleotide linkages are independently n001. In some embodiments, they are both chiral controlled and Rp. In some embodiments, the oligonucleotide comprises nucleoside N 0 wherein the nucleoside contains a natural DNA (two 2'-H), a natural RNA sugar, or a 2'-F modified sugar. 0 is the nucleoside opposite the target adenosine when the oligonucleotide is utilized for adenosine editing. 0 The sugars are natural DNA sugars. 1 (A "+" or nothing before the number indicates counting in the 5' direction (5'...N 1 N 0 N -1 ...3') sugar is a 2'-modified sugar, a natural DNA sugar, or a natural RNA sugar. 1In some embodiments, the sugar is a DNA sugar. -1 ("-" indicates counting in the 3' direction (5'...N 1 N 0 N -1 ...3') sugar is a 2'-F modified sugar, a natural DNA sugar, or a natural RNA sugar. -1 In some embodiments, the sugar is a DNA sugar. -3 In some embodiments, the sugar is a 2'-F modified sugar. 2 Between its 5' end, the oligonucleotide may contain a plurality of 2'-F modified sugars and a plurality of 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the oligonucleotide comprises a N 2 and at its 5' end, one or more (e.g., 1 to 20, 1 to 15, 1 to 10, 2 to 15, 2 to 10, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more) 2'-F blocks and one or more (e.g., 1 to 20, 1 to 15, 1 to 10, 2 to 15, 2 to 10, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more) separation blocks (e.g., 2 Ends with N 2where the first domain and the first subdomain of the second domain are linked, each nucleoside in the 2'-F block independently comprises a 2'-F modification and each nucleoside in the separation block independently does not comprise a 2'-F modification, and each block independently comprises one or more (e.g., 1-20, 1-15, 1-10, 2-15, 2-10, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more) nucleosides. In some embodiments, there are two or more such 2'-F blocks, and two or more such separation blocks. In some embodiments, one or more, or all, of such separation blocks are independently attached to two 2'-F blocks. In some embodiments, each nucleoside in one, more, or all of the separation blocks independently comprises a 2'-OR modification, where R is an optionally substituted C 1~6 In some embodiments, each nucleoside in one, more, or all of the separation blocks independently comprises a 2'-OR modification (wherein R is an optionally substituted C 1~6 In some embodiments, each nucleoside in one, more, or all of the separation blocks independently comprises a 2'-OMe or 2'-MOE modification. In some embodiments, each such 2'-F block and separation block independently comprises 1, 2, 3, 4, or 5 nucleosides. In some embodiments, N 0 Nearby nucleosides (e.g., N 2 , N 1 , N 0 , N -1 , N -2 etc.) do not contain bulky 2'-modifications such as 2'-MOE. 2 , N 1 , N 0 , N -1 , and N -2is independently a natural DNA sugar, a 2'-F modified sugar, or a 2'-OMe modified sugar. 1 , N 0 , N -1 each sugar is a natural DNA sugar. In some embodiments, each chiral internucleotide linkage is independently chiral controlled.

[0009] In some embodiments, the first domain comprises one or more 2'-F modifications and the second domain comprises one or more sugars that do not have a 2'-F modification. In some embodiments, the provided oligonucleotide comprises one or more chirally modified internucleotide linkages. In some embodiments, the first domain comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more sugars that comprise a 2'-F modification and a 2'-OR modification (wherein R is not -H) (e.g., 2-OMe, 2,-MOE, 2'-OL B -4'(in the formula, L B is an optionally substituted -CH 2 and 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more sugars, each independently comprising a 2'-OR modification (where R is not -H) (e.g., 2-OMe, 2,-MOE, 2'-OL B -4'(in the formula, L B is an optionally substituted -CH 2 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more sugars, each of which independently contains -, -, -, -;

[0010] In some embodiments, about 20%-80% (e.g., about 25%-80%, 30%-80%, 35%-80%, 40%-80%, 40%-70%, 40%-60%, 50%-80%, 50%-75%, 50%-60%, 55%-80%, 60-80%, or about 50%, 55%, 60%, 65%, 70%, 75%, or 80%) of the total sugars of the first domain comprise a 2'-F modification. In some embodiments, between about 20% and 70% (e.g., between about 20% and 60%, 20% and 50%, 30% and 60%, 30% and 50%, 40% and 50%, or about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60%) of the total sugars of the first domain are independently 2'-OR modifications (e.g., 2'-OMe, 2'-MOE, 2'-O-LB-4', where L B is an optionally substituted -CH 2 In some embodiments, the second domain comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more modified sugars that do not contain a 2'-F modification, or at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of all sugars in the second domain do not contain a 2'-F modification.

[0011] In some embodiments, the second domain comprises or consists of a first subdomain, a second subdomain, and a third subdomain as described herein. In some embodiments, the first subdomain comprises a 2'-OR modification (wherein R is not -H) (e.g., 2'-OMe, 2,-MOE, 2'-OL B -4'(in the formula, L B is an optionally substituted -CH 2 In some embodiments, the sugars in the second subdomain each independently comprise one or more (e.g., 1-10, 1-5, 1-3, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) sugars each independently comprising a 2'-OR modification (e.g., R is an optionally substituted C1~6 aliphatic), or 2'-OL B In some embodiments, each sugar of the second subdomain is independently a natural DNA sugar, a natural RNA sugar, or a 2'-F modified sugar. In some embodiments, each sugar of the second subdomain is independently a natural DNA sugar or a natural RNA sugar. In some embodiments, each sugar of the second subdomain is independently a natural DNA sugar or a 2'-F modified sugar. In some embodiments, each sugar of the second subdomain is independently a natural DNA sugar. In some embodiments, there are three nucleosides in the second subdomain. In some embodiments, these three of the second nucleosides are opposite the target adenosine when bound to the target. In some embodiments, the sugar of the second nucleoside does not contain any 2'-OR modifications (e.g., 2'-OMe, 2'-MOE, etc.) as described herein. In some embodiments, such sugar is a natural DNA sugar. In some embodiments, it is a natural RNA sugar. In some embodiments, this is a 2'-F modified sugar. In some embodiments, each third subdomain independently has a 2'-OR modification (wherein R is not -H) (e.g., 2'-OMe, 2,-MOE, 2'-OL B -4'(in the formula, L B is an optionally substituted -CH 2 -F modified sugars, etc.). In some embodiments, there are more such sugars in the third subdomain than there are 2'-F modified sugars.

[0012] In some embodiments, the second domain comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more modified sugars that independently comprise a 2'-OR modification, or at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of all sugars in the second domain are 2'-OR modified (wherein R is an optionally substituted C 1~6 In some embodiments, R is methyl. In some embodiments, R is -CH 2 CH 2 OCH 3 As described herein, other sugar modifications can also be utilized in accordance with the present disclosure, optionally in conjunction with the base modifications and / or internucleotide linkage modifications described herein.

[0013] In some embodiments, the oligonucleotide comprises or is a 5'-first domain-second domain-3' structure. In some embodiments, the second domain comprises or is a 5'-first subdomain-second subdomain-third subdomain-3' structure. In some embodiments, the oligonucleotide comprises or is a 5'-first domain-first subdomain-second subdomain-third subdomain-3' structure. In some embodiments, the oligonucleotide is conjugated to an additional moiety (e.g., various additional chemical moieties as described herein). In some embodiments, the oligonucleotide comprises an additional moiety (e.g., additional moieties as described herein). In some embodiments, the additional chemical moiety is or includes a small molecule moiety, a carbohydrate moiety (e.g., a GalNAc moiety), a nucleic acid moiety (e.g., an oligonucleotide moiety, a nucleic acid moiety, etc. (e.g., a moiety of an RNase H-dependent oligonucleotide, an RNAi oligonucleotide, an aptamer, a gRNA, etc.) that can provide and / or modulate one or more properties and / or activities), and / or a peptide moiety.

[0014] In some embodiments, the base sequence of the provided oligonucleotide is substantially complementary to the base sequence of the target nucleic acid, including the target adenosine. In some embodiments, the provided oligonucleotide contains one or more mismatches (non-Watson-Crick base pairs) when aligned with the target nucleic acid. In some embodiments, the provided oligonucleotide contains one or more wobble (e.g., GU, IA, GA, IU, IC, etc.) when aligned with the target nucleic acid. In some embodiments, the mismatch and / or wobble may aid one or more proteins (e.g., ADAR1, ADAR2, etc.) in recognizing the duplex formed by the provided oligonucleotide and the target nucleic acid. In some embodiments, the provided oligonucleotide forms a duplex with the target nucleic acid. In some embodiments, the ADAR protein recognizes and binds to such a duplex. In some embodiments, the nucleoside opposite the target adenosine is located in the middle of the provided oligonucleotide, for example, with 5-50 nucleosides on the 5' side and 1-50 nucleosides on the 3' side. In some embodiments, the 5' side has more nucleosides than the 3' side. In some embodiments, the 5' side has fewer nucleosides than the 3' side. In some embodiments, the 5' side has the same number of nucleosides as the 3' side.

[0015] In some embodiments, the identity, e.g., the identity of two nucleic acid sequences, is about or at least about 70%. In some embodiments, it is about or at least about 75%. In some embodiments, it is about or at least about 80%. In some embodiments, it is about or at least about 85%. In some embodiments, it is about or at least about 90%. In some embodiments, it is about or at least about 95%. In some embodiments, it is about 100%.

[0016] In some embodiments, by utilizing a variety of structural elements (e.g., a variety of modifications, stereochemistries, and patterns thereof), the present disclosure can achieve desirable properties and high activity with short oligonucleotides (e.g., about 20-40, 25-40, 25-35, 26-32, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 nucleobases in length).

[0017] In some embodiments, the provided oligonucleotide comprises a modified nucleobase. In some embodiments, the modified nucleobase promotes the modification of the target adenosine. In some embodiments, the nucleobase opposite the target adenine interacts less strongly with the target adenine (e.g., forms fewer hydrogen bonds) than U while maintaining interaction with the enzyme (e.g., ADAR) compared to when U is present. In some embodiments, the nucleobase opposite and / or its associated sugar provides some flexibility (e.g., compared to U) to the ability of the enzyme (e.g., ADAR1, ADAR2, etc.) to modify the target adenosine. In some embodiments, the nucleobase immediately 5' or 3' to the opposite nucleobase (relative to the target adenine; e.g., I and its derivatives) promotes the modification of the target adenine. In particular, the present disclosure recognizes that such nucleobases may provide less steric hindrance than G when the duplex of the provided oligonucleotide and its target nucleic acid interacts with a modifying enzyme (e.g., ADAR1 or ADAR2). In some embodiments, the base sequence of the oligonucleotide is selected (e.g., where multiple adenosine residues are suitable targets) and / or designed (e.g., through the use of various nucleobases described herein) such that steric hindrance can be reduced or eliminated (e.g., the adjacent nucleoside opposite target A is not G).

[0018] A variety of internucleotide linkages may be utilized in the oligonucleotides provided according to the present disclosure. In some embodiments, the oligonucleotide comprises one or more types of internucleotide linkages. In some embodiments, the oligonucleotide comprises two or more types of internucleotide linkages. In some embodiments, the oligonucleotide comprises at least three types of internucleotide linkages. In some embodiments, the linkage contains a linked phosphorus atom bound to an oxygen atom that is not bound to or part of the backbone sugar ("PO linkage", e.g., a natural phosphate linkage). In some embodiments, the linkage contains a linked phosphorus atom bound to a sulfur atom that is not bound to or part of the backbone sugar ("PS linkage", e.g., a phosphorothioate internucleotide linkage). In some embodiments, the linkage contains a linked phosphorus atom bound to a nitrogen atom that is not bound to or part of the backbone sugar ("PN linkage", e.g., n001). In some embodiments, the oligonucleotide comprises one or more PS linkages. In some embodiments, the oligonucleotide comprises one or more PO linkages. In some embodiments, the oligonucleotide comprises one or more PN linkages. In some embodiments, the oligonucleotide comprises one or more PS and one or more PO linkages. In some embodiments, the oligonucleotide comprises one or more PS and one or more PN linkages. In some embodiments, the oligonucleotide comprises one or more PS, one or more PN and one or more PO linkages.

[0019] In some embodiments, the first domain comprises one or more PO linkages, one or more PS linkages, and one or more PN linkages. In some embodiments, the first subdomain comprises one or more PO linkages, one or more PS linkages, and / or one or more PN linkages. In some embodiments, the first subdomain comprises one or more PO linkages. In some embodiments, the first subdomain comprises one or more natural phosphate linkages. In some embodiments, the second subdomain comprises one or more modified internucleotide linkages. In some embodiments, each internucleotide linkage attached to a nucleoside of the second subdomain is independently a modified internucleotide linkage. In some embodiments, each internucleotide linkage attached to a nucleoside of the second subdomain is independently a PS or PN linkage. In some embodiments, the third subdomain comprises one or more PO linkages, one or more PS linkages, and / or one or more PN linkages. In some embodiments, the third subdomain comprises one or more PO linkages. In some embodiments, the third subdomain comprises one or more natural phosphate linkages. In some embodiments, the third subdomain comprises one or more PS linkages. In some embodiments, the third subdomain comprises one or more PN linkages. In some embodiments, the third subdomain comprises one or more PO linkages, one or more PS linkages, and one or more PN linkages. In some embodiments, the first internucleotide linkage of the first domain or oligonucleotide is a PN linkage. In some embodiments, the last internucleotide linkage of the third subdomain or oligonucleotide is a PN linkage. In some embodiments, the native DNA sugar is linked to a modified internucleotide linkage. In some embodiments, the native DNA sugar is linked to a PN or PS internucleotide linkage. In some embodiments, each native DNA sugar in an oligonucleotide or portion thereof (e.g., the first domain, the first subdomain, the second subdomain, the third subdomain, etc.) is independently linked to a modified internucleotide linkage. In some embodiments, each native DNA sugar is independently linked to a PN or PS internucleotide linkage.In some embodiments, the natural RNA sugar is linked to a modified internucleotide linkage. In some embodiments, the natural RNA sugar is linked to a PN or PS internucleotide linkage. In some embodiments, each natural RNA sugar in an oligonucleotide or portion thereof (e.g., the first domain, the first subdomain, the second subdomain, the third subdomain, etc.) is independently linked to a modified internucleotide linkage. In some embodiments, each natural RNA sugar is independently linked to a PN or PS internucleotide linkage.

[0020] In some embodiments, the 2'-F modified sugar is linked to a modified internucleotide linkage. In some embodiments, the 2'-F modified sugar is linked to a PN or PS internucleotide linkage. In some embodiments, each 2'-F modified sugar in an oligonucleotide or portion thereof (e.g., the first domain, the first subdomain, the second subdomain, the third subdomain, etc.) is independently linked to a modified internucleotide linkage. In some embodiments, each 2'-F modified sugar is independently linked to a PN or PS internucleotide linkage. In some embodiments, each PO linkage is independently a natural phosphate linkage. In some embodiments, each PS linkage is independently a phosphorothioate internucleotide linkage. In some embodiments, one or more PN linkages are independently a non-negatively charged internucleotide linkage. In some embodiments, one or more PN linkages are independently a natural internucleotide linkage. In some embodiments, one or more PN linkages are independently a phosphorylguanidine linkage. In some embodiments, each PN linkage is independently a phosphorylguanidine linkage. In some embodiments, one or more PN bonds are independently n001. In some embodiments, each PN bond is independently n001.

[0021] In some embodiments, the oligonucleotides of the present disclosure provide modified internucleotide linkages (i.e., internucleotide linkages that are not natural phosphate linkages). In some embodiments, the linking phosphorus of the modified internucleotide linkages (e.g., chiral internucleotide linkages) is chiral and can exist in different configurations (Rp and Rs). In some embodiments, the incorporation of modified internucleotide linkages, particularly by controlling the stereochemistry of the linking phosphorus center (enriching one configuration at such controlled center compared to stereoirregular oligonucleotide preparations), can significantly improve properties (e.g., stability) and / or activity (e.g., adenosine modifying activity (e.g., converting adenosine to inosine)). In some embodiments, the oligonucleotides provided have significantly higher stereochemical purity compared to stereoirregular preparations. In some embodiments, the oligonucleotides provided are chiral controlled.

[0022] In some embodiments, oligonucleotides of the present disclosure include one or more chiral internucleotide linkages (e.g., phosphorothioate internucleotide linkages) in which the linked phosphorus is chiral. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% (e.g., 50%-100%, 60%-100% or 10 ... %, 70-100%, 75%-100%, 80%-100%, 90%-100%, 95%-100%, 60%-95%, 70%-95%, 75-95%, 80-95%, 85-95%, 90-95%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% (e.g., 70-100%, 75-100%, 80%-100%, 90%-100%, 95%-100%, 60%-95%, 70%-95%, 75-95%, 80-95%, 85-95%, 90-95%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% or all of the internucleotide linkages are chiral internucleotide linkages. In some embodiments, at least one internucleotide linkage is a chiral internucleotide linkage. In some embodiments, at least one internucleotide linkage is a natural phosphate linkage. In some embodiments, each internucleotide linkage is independently a chiral internucleotide linkage. In some embodiments, at least one chiral internucleotide linkage is a phosphorothioate internucleotide linkage. In some embodiments, each is a phosphorothioate internucleotide linkage. In some embodiments, one or more chiral internucleotide linkages are independently non-negatively charged or neutral internucleotide linkages. In some embodiments, one or more chiral internucleotide linkages are independently phosphorylguanidine internucleotide linkages. In some embodiments, one or more chiral internucleotide linkages are independently chiral controlled. In some embodiments, each chiral internucleotide linkage is independently chiral controlled. In some embodiments, one or more chiral internucleotide linkages are not chiral controlled. In some embodiments, each phosphorothioate internucleotide linkage is independently chiral controlled.In some embodiments, each modified internucleotide linkage is independently a phosphorothioate internucleotide linkage or a non-negatively charged internucleotide linkage. In some embodiments, each modified internucleotide linkage is independently a phosphorothioate internucleotide linkage or a neutral internucleotide linkage. In some embodiments, each modified internucleotide linkage is independently a phosphorothioate internucleotide linkage or a neutral internucleotide linkage. In some embodiments, each modified internucleotide linkage is independently a phosphorothioate internucleotide linkage or a phosphorylguanidine internucleotide linkage. In some embodiments, a phosphorylguanidine internucleotide linkage is n001. In some embodiments, a phosphorylguanidine internucleotide linkage is n001. In some embodiments, a non-negatively charged internucleotide linkage is n001. In some embodiments, a neutral internucleotide linkage is n001. In some embodiments, a modified internucleotide linkage is n002. In some embodiments, it is n006. In some embodiments, it is n020. In some embodiments, this is n004. In some embodiments, this is n008. In some embodiments, this is n025. In some embodiments, this is n026. A variety of modified internucleotide linkages are described herein. The bound phosphorus can be either Rp or Sp. In some embodiments, at least one bound phosphorus is Rp. In some embodiments, at least one bound phosphorus is Sp.In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% (e.g., between 50% and 100%, 60%, % to 100%, 70% to 100%, 75% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 60% to 95%, 70% to 95%, 75% to 95%, 80% to 95%, 85% to 95%, 90% to 95%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%, etc.), or all of the chiral internucleotide bonds are Sp. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% (e.g., between 50% and 100%, 60%, % to 100%, 70% to 100%, 75% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 60% to 95%, 70% to 95%, 75% to 95%, 80% to 95%, 85% to 95%, 90% to 95%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% (e.g., 100% to 100%, 70% to 100%, 75% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 60% to 95%, 70% to 95%, 75% to 95%, 80% to 95%, 85% to 95%, 90% to 95%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% or all phosphorothioate internucleotide linkages are Sp. In some embodiments, at least 50% of all phosphorothioate internucleotide linkages are Sp. In some embodiments, at least 60% of all phosphorothioate internucleotide linkages are Sp. In some embodiments, at least 70% of all phosphorothioate internucleotide linkages are Sp. In some embodiments, at least 75% of all phosphorothioate internucleotide linkages are Sp. In some embodiments, at least 80% of all phosphorothioate internucleotide linkages are Sp. In some embodiments, at least 85% of all phosphorothioate internucleotide linkages are Sp.In some embodiments, at least 90% of all phosphorothioate internucleotide linkages are Sp. In some embodiments, at least 95% of all phosphorothioate internucleotide linkages are Sp. In some embodiments, at least 96% of all phosphorothioate internucleotide linkages are Sp. In some embodiments, at least 97% of all phosphorothioate internucleotide linkages are Sp. In some embodiments, at least 98% of all phosphorothioate internucleotide linkages are Sp. In some embodiments, all phosphorothioate internucleotide linkages are Sp. In some embodiments, no more than 3, 4, 5, 6, 7, 8, 9, or 10 consecutive phosphorothioate internucleotide linkages are Rp. In some embodiments, no more than 3 consecutive phosphorothioate internucleotide linkages are Rp. In some embodiments, no more than 4 consecutive phosphorothioate internucleotide linkages are Rp. In some embodiments, no more than 5 consecutive phosphorothioate internucleotide linkages are Rp. In some embodiments, no more than 6 consecutive phosphorothioate internucleotide linkages are Rp. In some embodiments, no more than 7 consecutive phosphorothioate internucleotide linkages are Rp. In some embodiments, no more than 8 consecutive phosphorothioate internucleotide linkages are Rp. In some embodiments, no more than 9 consecutive phosphorothioate internucleotide linkages are Rp. In some embodiments, no more than 10 consecutive phosphorothioate internucleotide linkages are Rp. In some embodiments, no consecutive Rp phosphorothioate internucleotide linkages are utilized in moieties where a majority (e.g., greater than 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95% or higher) or all of the sugars are naturally-occurring DNA, and / or RNA, and / or 2'-F modified sugars.In some embodiments, when phosphorothioate internucleotide linkages of consecutive Rp are utilized, one or more or a majority (e.g., greater than 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95% or higher), or all of such internucleotide linkages are independently linked to a sugar that may improve stability. In some embodiments, when phosphorothioate internucleotide linkages of consecutive Rp are utilized, one or more or a majority (e.g., greater than 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95% or higher), or all of such internucleotide linkages are independently linked to a bicyclic sugar or a 2'-OR modified sugar (wherein R is an optionally substituted C. 1~6 In some embodiments, when phosphorothioate internucleotide linkages of consecutive Rp are utilized, one or more or a majority (e.g., greater than 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95% or higher), or all of such internucleotide linkages are independently linked to a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 aliphatic). In some embodiments, each 2'-OR modified sugar is independently a 2'-OMe modified sugar or a 2'-MOE modified sugar. In some embodiments, each 2'-OR modified sugar is independently a 2'-OMe modified sugar. In some embodiments, each 2'-OR modified sugar is independently a 2'-MOE modified sugar.

[0023] In some embodiments, the stereochemistry of one or more chiral linkage phosphorus of a provided oligonucleotide is controlled in the composition. In some embodiments, the disclosure provides a composition comprising a plurality of oligonucleotides, the plurality of oligonucleotides having a common base sequence and one or more (e.g., about 1-50, 1-40, 1-30, 1-25, 1-20, 1-15, 1-10, 5-50, 5-40, 5-30, 5-25, 5-20, 5-15, 5-10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 1 2, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 or more, or at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of the chiral internucleotide linkages ("chiral controlled internucleotide linkages") share the same configuration of the independent linking phosphorus (e.g., all Rp or all Sp with respect to the chiral linking phosphorus). In some embodiments, they share the same stereochemistry at their respective chiral linking phosphorus. In some embodiments, the multiple oligonucleotides share the same configuration. In some embodiments, the multiple oligonucleotides are structurally identical except for the internucleotide linkage. In some embodiments, the multiple oligonucleotides are structurally identical. In some embodiments, at least about 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of all of the oligonucleotides in the composition, or all of the oligonucleotides that share a common base sequence, share the pattern of chiral centers in the backbone of the plurality of oligonucleotides. In some embodiments, at least about 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of all of the oligonucleotides in the composition, or all of the oligonucleotides that share a common base sequence, are a plurality of oligonucleotides.

[0024] In some embodiments, the disclosure provides chiral controlled oligonucleotide compositions of oligonucleotides, wherein at least about 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of all oligonucleotides in the composition, or all oligonucleotides having the same base sequence, or all oligonucleotides having the same base sequence and sugar and base modifications, or all oligonucleotides of the same composition, have one or more (e.g., about 1-50, 1-4, or 50% of all chiral internucleotide linkages) with the oligonucleotide. 0, 1 to 30, 1 to 25, 1 to 20, 1 to 15, 1 to 10, 5 to 50, 5 to 40, 5 to 30, 5 to 25, 5 to 20, 5 to 15, 5 to 10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 or more, or at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%, of the chiral internucleotide bonds independently share the same configuration of bonding phosphorus (e.g., all of the chiral bonding phosphorus are Rp or all of the chiral bonding phosphorus are Sp). In some embodiments, the disclosure provides chiral controlled oligonucleotide compositions of oligonucleotides, wherein at least about 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of all of the oligonucleotides in the composition, or all of the oligonucleotides having the same base sequence, or all of the oligonucleotides having the same base sequence and sugar and base modifications, or all of the oligonucleotides of the same composition, are in one or more forms of the oligonucleotide (e.g., an acid form, a salt form (e.g., a pharma- ceutically acceptable salt form; as will be understood by one of skill in the art, if the oligonucleotide is a salt, other salt forms of the corresponding acid or base form of the oligonucleotide), etc.).

[0025] In some embodiments, chiral controlled oligonucleotide compositions provide several advantages (e.g., higher stability, activity, etc.) compared to corresponding stereoirregular oligonucleotide compositions. In some embodiments, chiral controlled oligonucleotide compositions provide high levels of adenosine modifying (e.g., A to I conversion) activity at various isoforms of ADAR proteins (e.g., p150 and p110 forms of ADAR1), while corresponding stereoirregular compositions provide high levels of adenosine modifying (e.g., A to I conversion) activity at only certain isoforms of ADAR proteins (e.g., p150 isoform of ADAR1).

[0026] In some embodiments, provided oligonucleotides include an additional moiety (e.g., a targeting moiety, a carbohydrate moiety, etc.). In some embodiments, the additional moiety is or includes a ligand for the asialoglycoprotein receptor. In some embodiments, the additional moiety is or includes GalNAc or a derivative thereof. In particular, the additional moiety may facilitate delivery to a particular target site (e.g., a cell, tissue, organ, etc. (e.g., a site that includes a receptor that interacts with the additional moiety)). In some embodiments, the additional moiety facilitates delivery to the liver.

[0027] In some embodiments, the present disclosure provides techniques for preparing oligonucleotides and compositions thereof, particularly chiral controlled oligonucleotide compositions. In some embodiments, the oligonucleotides and compositions thereof provided are of high purity. In some embodiments, the oligonucleotides of the present disclosure are at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% stereochemically pure at the linkage phosphorus of the chiral internucleotide linkage. In some embodiments, the oligonucleotides of the present disclosure are stereoselectively prepared and are substantially free of stereoisomers. In some embodiments, in compositions provided that include multiple oligonucleotides sharing the same base sequence with the same pattern of stereochemistry of the chiral bond phosphorus (e.g., each chiral bond phosphorus includes one or more of Rp and / or Sp, independently Rp or Sp), at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of all oligonucleotides in the composition that share the same base sequence with the multiple oligonucleotides share the same pattern of stereochemistry of the chiral bond phosphorus, or are the multiple oligonucleotides. In some embodiments, in compositions provided that include multiple oligonucleotides sharing the same base sequence with the same pattern of stereochemistry of the chiral bond phosphorus, at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of all oligonucleotides in the composition that share the same configuration with the multiple oligonucleotides share the same pattern of stereochemistry of the chiral bond phosphorus, or are the multiple oligonucleotides.

[0028] In some embodiments, the present disclosure describes useful techniques for evaluating oligonucleotides and compositions thereof. For example, various techniques of the present disclosure are useful for evaluating adenosine modification. As will be understood by those skilled in the art, in some embodiments, adenosine modification / editing can be evaluated via sequencing, mass spectrometry, evaluation (e.g., levels, activity, etc.) of products (e.g., RNA, protein, etc.) of modified nucleic acid (e.g., adenosine of target nucleic acid is converted to inosine), optionally in view of the presence of other components (e.g., ADAR protein) in the modification system (e.g., in vitro system, ex vivo system, cell, tissue, organ, organism, subject, etc.). Those skilled in the art will understand that an oligonucleotide that results in adenosine modification of a target nucleic acid can also provide a modified nucleic acid (e.g., adenosine of target nucleic acid is converted to I) and one or more of its products (e.g., mRNA, protein, etc.). Certain useful techniques are described in the examples.

[0029] As described herein, the oligonucleotides and compositions of the disclosure may be provided / utilized in a variety of forms. In some embodiments, the disclosure provides for the oligonucleotides to be used in one or more forms (e.g., an acid form (e.g., a native phosphate linkage exists as -O(P(O)(OH)-O- and a phosphorothioate internucleotide linkage exists as -O(P(O)(SH)-O-), a base form, a salt form (e.g., a native phosphate linkage exists as a salt form (e.g., a sodium salt (-O(P(O)(OH)-O-)), a phosphate linkage exists as ... - Na + )-O-), and phosphorothioate internucleotide linkages exist in salt form (e.g., sodium salt (-O(P(O)(S - Na +As will be appreciated by those skilled in the art, oligonucleotides may exist in various salt forms, including pharma- ceutically acceptable salts, and in solutions (e.g., various aqueous buffer systems), in which the cation may dissociate from the anion. In some embodiments, the present disclosure provides a pharmaceutical composition comprising the provided oligonucleotide and / or one or more pharma-ceutically acceptable salts thereof, and a pharma-ceutically acceptable carrier. In some embodiments, the pharmaceutical composition is a chiral controlled oligonucleotide composition.

[0030] The provided technology can be utilized for a variety of purposes. For example, one of skill in the art will appreciate that the provided technology is useful for many purposes, including modifying adenosines, e.g., correcting G to A mutations, modulating the levels of certain nucleic acids and / or products encoded thereby (e.g., reducing protein levels by introducing A to G / I modifications), modulating splicing, modulating translation (e.g., modulating translation start and / or stop sites by introducing A to G / I modifications), and the like.

[0031] In some embodiments, the disclosure provides techniques for preventing or treating conditions, disorders, or diseases that are susceptible to adenosine modifications (e.g., A to I or G conversion). As will be appreciated by those skilled in the art, I may perform one or more functions of G, for example, in base pairing, translation, etc. In some embodiments, the G to A mutation may be corrected via A to I conversion so that one or more products (e.g., proteins) of the G form of the nucleic acid may be generated. In some embodiments, the disclosure provides techniques for preventing or treating conditions, disorders, or diseases associated with the mutation, comprising administering to a subject susceptible to or suffering from the same a provided oligonucleotide or composition thereof, where the oligonucleotide or composition may edit the mutation. In some embodiments, the disclosure provides techniques for preventing or treating conditions, disorders, or diseases associated with the G to A mutation, comprising administering to a subject susceptible to or suffering from the same a provided oligonucleotide or composition thereof, where the oligonucleotide or composition may modify the A. In some embodiments, the provided technology modifies A in a transcript (e.g., an RNA transcript). In some embodiments, A is converted to I. In some embodiments, during translation, the protein synthesis machinery reads I as G. In some embodiments, a G / I type has or encodes one or more proteins that have one or more higher desired activities and / or one or more better desired properties compared to its corresponding A type and / or one or more proteins encoded thereby. In some embodiments, a G / I type provides higher levels of one or more proteins that have one or more higher desired activities and / or one or more better desired properties compared to its corresponding A type. In some embodiments, a product encoded by a G / I type is structurally different (e.g., a longer, in some embodiments, full-length protein) than that encoded by its corresponding A type. In some embodiments, a G / I type provides a structurally identical product (e.g., a protein) compared to its corresponding A type, but the G / I type provides such a product at a more desirable level.

[0032] As one of skill in the art will appreciate, many conditions, disorders, or diseases are associated with mutations that can be altered by the provided techniques and can be prevented and / or treated using the provided techniques. For example, over 20,000 conditions, disorders, or diseases have been reported to be associated with G to A mutations and could benefit from A to I editing. [Brief description of the drawings]

[0033] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1-1] The provided technology can provide editing. Various oligonucleotides were evaluated, including first and second domains, where the corresponding first and second portions are separated by gaps of various lengths (in some embodiments, 1, 3, 5, 10, 30, 60, 137, 200, or 206 nucleobases). As shown in (A) and (B), various oligonucleotides can provide editing of the target adenosine. The oligonucleotides were transfected into 293T cells expressing ADAR-110 (A) or ADAR-150 (B). [Figure 1-2] The provided technology can provide editing. Various oligonucleotides were evaluated, including first and second domains, where the corresponding first and second portions are separated by gaps of various lengths (in some embodiments, 1, 3, 5, 10, 30, 60, 137, 200, or 206 nucleobases). As shown in (A) and (B), various oligonucleotides can provide editing of the target adenosine. The oligonucleotides were transfected into 293T cells expressing ADAR-110 (A) or ADAR-150 (B). [Diagram 2]The provided technology can provide editing. Various oligonucleotides were evaluated, including a first domain and a second domain, with corresponding first and second portions in the NRF2 transcript separated by gaps of various lengths. Various oligonucleotides can provide editing of target adenosines in the NRF2 transcript. Oligonucleotides were transfected into human primary astrocytes expressing endogenous ADAR. Gap length is indicated in brackets. Normal = no gap. Error bars represent standard error of the mean (SEM), N=2. [Diagram 3] The provided technology can provide editing. Various oligonucleotides, including a first domain complementary to a first target adenosine and a second domain complementary to a second target adenosine, can provide editing of a target adenosine in NRF2 transcript. The oligonucleotides were transfected into human primary astrocytes expressing endogenous ADAR. The target adenosine is referenced by the relevant amino acid codon (e.g., Q26 indicates editing of an adenosine in a glutamine codon corresponding to position 26 of NRF2 protein). The position of the nucleoside opposite the target adenosine is indicated in brackets (e.g., P10 indicates nucleoside 10 of the oligonucleotide when counting from 5' to 3'). Error bars represent standard error of the mean (SEM), N=2. [Figure 4] The provided technology can provide editing. Various oligonucleotides were evaluated, including a first domain and a second domain, with the corresponding first and second portions separated by gaps of various lengths. Various oligonucleotides can provide editing of the target adenosine within the Malat1 transcript. For exogenous ADAR expression, a plasmid encoding ADAR1 p110 was transfected into HEK293T cells. Oligonucleotides were transfected into cells at a dose of 25 nM after 24 hours. Gap length is indicated in brackets. Standard = no gap. Error bars represent standard error of the mean (SEM), N=2. [Figure 5-1]The provided technology can provide editing. Various oligonucleotides were evaluated, including a first domain and a second domain, with the corresponding first and second portions separated by gaps of various lengths. As shown in (A), (B), and (C), various oligonucleotides can provide editing of the target adenosine in the MECP2 transcript. HEK293T cells were transfected with a plasmid encoding ADAR1 p110 for exogenous ADAR expression, and a plasmid encoding MECP2 for exogenous MECP2 expression. Oligonucleotides were transfected into cells at a dose of 25 nM after 24 hours. Gap lengths are indicated in brackets. Error bars represent standard error of the mean (SEM), N=2. [Figure 5-2] The provided technology can provide editing. Various oligonucleotides were evaluated, including a first domain and a second domain, with the corresponding first and second portions separated by gaps of various lengths. As shown in (A), (B), and (C), various oligonucleotides can provide editing of the target adenosine in the MECP2 transcript. HEK293T cells were transfected with a plasmid encoding ADAR1 p110 for exogenous ADAR expression, and a plasmid encoding MECP2 for exogenous MECP2 expression. Oligonucleotides were transfected into cells at a dose of 25 nM after 24 hours. Gap lengths are indicated in brackets. Error bars represent standard error of the mean (SEM), N=2. [Figure 5-3]The provided technology can provide editing. Various oligonucleotides were evaluated, including a first domain and a second domain, with the corresponding first and second portions separated by gaps of various lengths. As shown in (A), (B), and (C), various oligonucleotides can provide editing of the target adenosine in the MECP2 transcript. HEK293T cells were transfected with a plasmid encoding ADAR1 p110 for exogenous ADAR expression, and a plasmid encoding MECP2 for exogenous MECP2 expression. Oligonucleotides were transfected into cells at a dose of 25 nM after 24 hours. Gap lengths are indicated in brackets. Error bars represent standard error of the mean (SEM), N=2. [Figure 6] The provided technology can provide editing. Various oligonucleotides were evaluated, including a first domain and a second domain, with the corresponding first and second portions separated by gaps of various lengths. Oligonucleotides including various linkers (e.g., [SpC3], [oC4o], [oC6o], [Sp9], [Sp12]) can provide editing of the target adenosine in the MECP2 transcript. HEK293T cells were transfected with a plasmid encoding ADAR1 p110 for exogenous ADAR expression, and a plasmid encoding MECP2 for exogenous MECP2 expression. Oligonucleotides were transfected into cells at a dose of 25 nM after 24 hours. The gap length of the oligonucleotides is indicated at the bottom. Error bars represent standard error of the mean (SEM), N=2. [Figure 7]The provided technology can provide editing. Various oligonucleotides were evaluated, including a first domain and a second domain, with the corresponding first and second portions separated by gaps of various lengths. Oligonucleotides containing various linkers (e.g., [SpC3], [oC4o], [oC6o], [Sp9], [Sp12]) can provide editing of the target adenosine in the Malat1 transcript. For exogenous ADAR expression, a plasmid encoding ADAR1 p110 was transfected into HEK293T cells. Oligonucleotides were transfected into cells at a dose of 25 nM after 24 hours. The gap length of the oligonucleotides is indicated at the bottom. Error bars represent standard error of the mean (SEM), N=2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] Detailed Description of Specific Embodiments The techniques of the present disclosure may be more readily understood by reference to the following detailed description of specific embodiments.

[0035] definition As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this disclosure, chemical elements are identified according to the Periodic Table of the Elements, CAS system, Handbook of Chemistry and Physics, 75th Ed. Furthermore, general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999 and "March's Advanced Organic Chemistry", 5th Ed., Ed.: Smith, M. B. and March, J., John Wiley & Sons, New York: 2001.

[0036] As used herein in this disclosure, unless otherwise clear from the context, (i) the term "a" or "an" may be understood to mean "at least one"; (ii) the term "or" may be understood to mean "and / or"; (iii) the terms "comprising," "comprise," "including" (whether or not used in conjunction with "not limited to"), and "include" (whether or not used in conjunction with "not limited to") may be understood to encompass the itemized components or steps, whether presented by themselves or with one or more additional components or steps; (iv) the term "another" may be understood to mean at least an additional / second one or more; (v) the terms "about" and "approximately" may be understood to allow for standard variations as understood by one of ordinary skill in the art; (vi) when ranges are given, the endpoints are included.

[0037] Unless otherwise specified, descriptions of oligonucleotides and their elements (e.g., base sequence, sugar modifications, internucleotide bonds, stereochemistry of the linking phosphorus, patterns thereof, etc.) are in the 5' to 3' direction. As one of skill in the art will appreciate, in some embodiments, oligonucleotides may be provided and / or utilized as salt forms, particularly pharma- ceutically acceptable salt forms, such as sodium salts. As one of skill in the art will also appreciate, in some embodiments, individual oligonucleotides within a composition may be considered to be of the same composition and / or structure even within such a composition (e.g., a liquid composition), and a particular such oligonucleotide may be in different salt forms at a particular moment (and may be dissolved or may exist as an anionic form when the oligonucleotide chain is, for example, in a liquid composition). For example, one of skill in the art will appreciate that at a given pH, individual internucleotide bonds along an oligonucleotide chain may be in the acid (H) form or one of several possible salt forms (e.g., sodium salts, or salts of different cations depending on the ions that may be present in the preparation or composition), and may be in their acid form (e.g., all cations, if present, are H salts). + It will be understood that so long as the first and second oligonucleotides (replaced with ) are of the same composition and / or structure, such individual oligonucleotides may be considered to be of the same composition and / or structure, as appropriate.

[0038] Aliphatic: As used herein, "aliphatic" refers to a linear (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain containing one or more units that are fully saturated or unsaturated (but not aromatic), or a substituted or unsubstituted monocyclic, bicyclic, or polycyclic hydrocarbon ring that is fully saturated or contains one or more units that are unsaturated (but not aromatic), or a combination thereof. In some embodiments, an aliphatic group contains 1-50 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-20 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-10 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-9 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-8 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-7 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-6 aliphatic carbon atoms. In still other embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms, and in yet other embodiments, aliphatic groups contain 1, 2, 3, or 4 aliphatic carbon atoms. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups, and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

[0039] Alkenyl: As used herein, the term "alkenyl" refers to an aliphatic group, as defined herein, having one or more double bonds.

[0040] Alkyl: As used herein, the term "alkyl" has its ordinary meaning in the art and can include saturated aliphatic groups, including straight chain alkyl groups, branched chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl groups, and cycloalkyl substituted alkyl groups. In some embodiments, an alkyl has 1-100 carbon atoms. In certain embodiments, a straight or branched chain alkyl has about 1-20 carbon atoms in its backbone (e.g., C for a straight chain). 1 ~C 20, C for branched chains 2 ~C 20 ), or about 1-10. In some embodiments, cycloalkyl rings have from about 3-10 carbon atoms in their ring structure, and such rings are monocyclic, bicyclic or polycyclic, or have about 5, 6 or 7 carbons in the ring structure. In some embodiments, an alkyl group can be a lower alkyl group, where a lower alkyl group contains 1-4 carbon atoms (e.g., C for a straight chain lower alkyl). 1 ~C 4 ).

[0041] Alkynyl: As used herein, the term "alkynyl" refers to an aliphatic group, as defined herein, having one or more triple bonds.

[0042] Analog: The term "analog" includes any chemical moiety that is structurally distinct from a reference chemical moiety or class of moieties, but can perform at least one function of such reference chemical moiety or class of moieties. Non-limiting examples include a nucleotide analog that is structurally distinct from a nucleotide, but performs at least one function of a nucleotide, a nucleobase analog that is structurally distinct from a nucleobase, but performs at least one function of a nucleobase, etc.

[0043] Animal: As used herein, the term "animal" refers to any member of the animal kingdom. In some embodiments, "animal" refers to humans at any stage of development. In some embodiments, "animal" refers to non-human animals at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., a rodent, mouse, rat, rabbit, monkey, dog, cat, sheep, cow, primate, and / or pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, and / or parasites. In some embodiments, the animal may be a transgenic animal, a genetically engineered animal, and / or a clone.

[0044] Aryl: The term "aryl," used alone or as part of a larger moiety of "aralkyl," "aralkoxy," or "aryloxyalkyl," as used herein, refers to a monocyclic, bicyclic, or polycyclic ring system having a total of 5-30 ring members, where at least one ring in these systems is aromatic. In some embodiments, the aryl group is a monocyclic, bicyclic, or polycyclic ring system having a total of 5-14 ring members, where at least one ring in these systems is aromatic, where each ring in these systems contains 3-7 ring members. In some embodiments, each monocyclic ring unit is aromatic. In some embodiments, the aryl group is a biaryl group. The term "aryl" may be used interchangeably with the term "aryl ring." In certain embodiments of the present disclosure, "aryl" refers to an aromatic ring system, including, but not limited to, phenyl, biphenyl, naphthyl, binaphthyl, anthracyl, and the like, which may bear one or more substituents. As used herein, also included within the scope of the term "aryl" are groups in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthymidyl, phenanthridinyl, or tetrahydronaphthyl.

[0045] Characteristic portion: As used herein, the term "characteristic portion" in its broadest sense refers to a portion of a substance whose presence (or absence) correlates with the presence (or absence) of a particular characteristic, attribute, or activity of the substance. In some embodiments, a characteristic portion of a substance is a portion found in the substance and in related substances that share the particular characteristic, attribute, or activity, but not in those that do not share the particular characteristic, attribute, or activity. In certain embodiments, a characteristic portion shares at least one functional characteristic with the intact substance. For example, in some embodiments, a "characteristic portion" of a protein or polypeptide is one that contains a stretch of contiguous amino acids, or a collection of stretches of contiguous amino acids, that together are characteristic of the protein or polypeptide. In some embodiments, each such contiguous stretch generally contains at least 2, 5, 10, 15, 20, 50 or more amino acids. Generally, a characteristic portion of a substance (e.g., a protein, antibody, etc.) is one that shares at least one functional characteristic with the related intact substance in addition to the sequence and / or structural identity specified above. In some embodiments, a characteristic portion may be biologically active.

[0046] Chiral control: As used herein, "chiral control" refers to the control of the stereochemical assignment of the chiral linkage phosphorus at the chiral internucleotide linkage in an oligonucleotide. As used herein, a chiral internucleotide linkage is an internucleotide linkage in which the linkage phosphorus is chiral. In some embodiments, the control is achieved by a chiral element not present in the sugar and base moieties of the oligonucleotide, for example, in some embodiments, the control is achieved by the use of one or more chiral auxiliary agents during oligonucleotide preparation, which are often part of the chiral phosphoramidite used during oligonucleotide preparation. In contrast to chiral control, those skilled in the art will understand that conventional oligonucleotide synthesis without the use of chiral auxiliary agents cannot control the stereochemistry at the chiral internucleotide linkage when such conventional oligonucleotide synthesis is used to form the chiral internucleotide linkage. In some embodiments, the stereochemical assignment of each chiral linkage phosphorus at each chiral internucleotide linkage in an oligonucleotide is controlled.

[0047] Chiral controlled oligonucleotide composition: The terms "chiral controlled oligonucleotide composition", "chiral controlled nucleic acid composition" and the like, as used herein, refer to a composition comprising multiple oligonucleotides (or nucleic acids) that share a common base sequence, and the multiple oligonucleotides (or nucleic acids) share the same linking phosphorus stereochemistry at one or more chiral internucleotide linkages (chiral controlled or sterically restricted internucleotide linkages, where the chiral linking phosphorus in the composition is Rp or Sp ("sterically restricted"), rather than a random mixture of Rp and Sp as non-chirally controlled internucleotide linkages). In some embodiments, a chiral controlled oligonucleotide composition comprises a plurality of oligonucleotides (or nucleic acids) that share 1) a common base sequence, 2) a common pattern of backbone linkages, and 3) a common pattern of backbone phosphorus modifications, where the plurality of oligonucleotides (or nucleic acids) share the same linkage phosphorus stereochemistry at one or more chiral internucleotide linkages (chiral controlled or sterically restricted internucleotide linkages, where the chiral linkage phosphorus is Rp or Sp in the composition ("sterically restricted"), rather than a random Rp and Sp mixture as a non-chiral controlled internucleotide linkage). The level of the plurality of oligonucleotides (or nucleic acids) in the chiral controlled oligonucleotide composition is predefined / controlled or enhanced (e.g., by a chiral controlled oligonucleotide preparation that stereoselectively forms one or more chiral internucleotide linkages).In some embodiments, about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 50%, ... 95%, 50%, 50%, 60%, 70%, 80%, 90%, 95%, 50%, 50%, 60%, 70%, 80%, 90%, 95%, 50%, 50%, 50%, 60%, 70%, 80%, 80%, 90%, 95%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, %, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%) are a plurality of oligonucleotides. In some embodiments, about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95 ...95% to 100%, 95% to 100%, 90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%) are a plurality of oligonucleotides.In some embodiments, the level is between about 1% and 100% (e.g., between about 5% and 100%) of all oligonucleotides in a composition, or of all oligonucleotides in a composition (e.g., of multiple oligonucleotides or types of oligonucleotides) that share a common base sequence, a common backbone linkage pattern, and a common backbone phosphorus modification pattern, or of all oligonucleotides in a composition that share a common base sequence, a common base modification pattern, a common sugar modification pattern, a common internucleotide linkage type pattern, and / or a common internucleotide linkage modification pattern. %, 10%-100%, 20%-100%, 30%-100%, 40%-100%, 50%-100%, 60%-100%, 70%-100%, 80-100%, 90-100%, 95-100%, 50%-90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 9 0%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%). In some embodiments, the oligonucleotides share the same stereochemistry at about 1-50 (e.g., about 1-10, 1-20, 5-10, 5-20, 10-15, 10-20, 10-25, 10-30, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) chiral internucleotide linkages.In some embodiments, the plurality of oligonucleotides comprises between about 1% and 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of the oligonucleotides (or nucleic acids) share the same stereochemistry. In some embodiments, multiple oligonucleotides (or nucleic acids) share the same pattern of sugar and / or nucleobase modifications in any one of the oligonucleotides. In some embodiments, multiple oligonucleotides (or nucleic acids) are various forms of the same oligonucleotide (e.g., acids and / or various salts of the same oligonucleotide). In some embodiments, multiple oligonucleotides (or nucleic acids) are of the same composition. In some embodiments, the level of the plurality of oligonucleotides (or nucleic acids) is between about 1% and 100% (e.g., between about 5% and 100%, 10% and 100%, 20% and 100%, 30% and 100%, 40% and 100%, 50% and 100%, 60% and 100%, 70% and 100%, 80% and 100%, 90% and 100%, 95% and 100% of all oligonucleotides (or nucleic acids) in the composition that share the same composition as the plurality of oligonucleotides (or nucleic acids). %, 50%-90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.In some embodiments, each chiral internucleotide linkage is a chiral controlled internucleotide linkage, and the composition is a completely chiral controlled oligonucleotide composition. In some embodiments, the multiple oligonucleotides (or nucleic acids) are structurally identical. In some embodiments, the chiral controlled internucleotide linkage has a diastereomeric purity of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5%, typically at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5%. In some embodiments, the chiral controlled internucleotide linkage has a diastereomeric purity of at least 95%. In some embodiments, the chiral controlled internucleotide linkage has a diastereomeric purity of at least 96%. In some embodiments, the chiral controlled internucleotide linkage has a diastereomeric purity of at least 97%. In some embodiments, the chiral controlled internucleotide linkage has a diastereomeric purity of at least 98%. In some embodiments, the chiral controlled internucleotide linkage has a diastereomeric purity of at least 99%. In some embodiments, the percentage of the level is (DS). nc or at least (DS) nc where DS is the diastereomeric purity as described herein (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% or more), and nc is the number of chiral controlled internucleotide linkages as described herein (e.g., 1-50, 1-40, 1-30, 1-25, 1-20, 5-50, 5-40, 5-30, 5-25, 5-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more). In some embodiments, the percentage level is (DS) nc or at least (DS) ncand DS is 95% to 100%. For example, when DS is 99% and nc is 10, the percentage is 90% or at least 90% ((99%) 10≈0.90=90%). In some embodiments, the level of the plurality of oligonucleotides in the composition is expressed as the product of the diastereopurity of each chiral controlled internucleotide bond in the oligonucleotide. In some embodiments, the diastereopurity of an internucleotide bond linking two nucleosides in an oligonucleotide (or nucleic acid) is expressed by the diastereopurity of the internucleotide bond of a dimer linking the same two nucleosides, where the dimer is prepared using equivalent conditions, in some examples, identical synthesis cycle conditions (e.g., in a bond between Nx and Ny in an oligonucleotide....NxNy....., the dimer is NxNy). In some embodiments, not all chiral internucleotide bonds are chiral controlled internucleotide bonds, and the composition is a partially chiral controlled oligonucleotide composition. In some embodiments, the non-chirally controlled internucleotide linkages have a diastereomeric purity of less than about 80%, 75%, 70%, 65%, 60%, 55%, or about 50%, as typically observed in stereorandom oligonucleotide compositions (e.g., from conventional oligonucleotide synthesis, e.g., phosphoramidite methods, as will be appreciated by those of skill in the art). In some embodiments, the multiple oligonucleotides (or nucleic acids) are of the same type. In some embodiments, the chiral controlled oligonucleotide composition comprises non-random or controlled levels of individual oligonucleotide or nucleic acid types. For example, in some embodiments, the chiral controlled oligonucleotide composition comprises only one oligonucleotide type. In some embodiments, the chiral controlled oligonucleotide composition comprises two or more oligonucleotide types. In some embodiments, the chiral controlled oligonucleotide composition comprises multiple oligonucleotide types. In some embodiments, the chiral controlled oligonucleotide composition is a composition of oligonucleotides of an oligonucleotide type, which composition comprises non-random or controlled levels of that oligonucleotide type. The method comprises the steps of:

[0048] Equivalent: The term "equivalent" is used herein to describe two (or more) sets of conditions or circumstances that are sufficiently similar to each other to allow for a comparison of the results obtained or the events observed. In some embodiments, comparable sets of conditions or circumstances are characterized by a number of substantially equivalent characteristics and one or a few varying characteristics. One skilled in the art will understand that sets of conditions are comparable to each other when they are characterized by a sufficient number and variety of substantially identical characteristics to warrant a reasonable conclusion that differences in the results obtained or the events observed under the different sets of conditions or circumstances are due to or imply changes in those characteristics that have been altered.

[0049] Alicyclic: The terms "alicyclic", "carbocycle", "carbocyclyl", "carbocyclic group", and "carbocyclic ring" are used interchangeably and as used herein, unless otherwise specified, refer to a saturated or partially unsaturated but non-aromatic cycloaliphatic monocyclic, bicyclic, or polycyclic ring system as described herein having 3 to 30 ring members. Alicyclic groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, norbornyl, adamantyl, and cyclooctadienyl. In some embodiments, an alicyclic group has 3 to 6 carbons. In some embodiments, an alicyclic group is saturated and is cycloalkyl. The term "alicyclic" may also include alicyclic rings fused to one or more aromatic or non-aromatic rings, such as decahydronaphthyl or tetrahydronaphthyl. In some embodiments, alicyclic groups are bicyclic. In some embodiments, alicyclic groups are tricyclic. In some embodiments, alicyclic groups are polycyclic. In some embodiments, "alicyclic" refers to a C ring having a single point of attachment to the remainder of the molecule that is fully saturated or contains one or more units of unsaturation, but is not aromatic. 3 ~C 6 Monocyclic hydrocarbons, or C 8 ~C10 Bicyclic or polycyclic hydrocarbons, or C, having a single point of attachment to the rest of the molecule that is fully saturated or contains one or more units of unsaturation but is not aromatic 9 ~C 16 Refers to polycyclic hydrocarbons.

[0050] Heteroaliphatic: The term "heteroaliphatic," as used herein, has its ordinary meaning in the art and refers to an aliphatic group, as described herein, in which one or more carbon atoms are independently replaced with one or more heteroatoms (e.g., oxygen, nitrogen, sulfur, silicon, phosphorus, etc.). In some embodiments, C, CH, CH 2 , and C.H. 3 are independently replaced by one or more heteroatoms (including oxidized and / or substituted forms thereof). In some embodiments, a heteroaliphatic group is a heteroalkyl. In some embodiments, a heteroaliphatic group is a heteroalkenyl.

[0051] Heteroalkyl: The term "heteroalkyl," as used herein, has its ordinary meaning in the art and refers to an alkyl group, as described herein, in which one or more carbon atoms are independently replaced with one or more heteroatoms (e.g., oxygen, nitrogen, sulfur, silicon, phosphorus, etc.). Examples of heteroalkyl include, but are not limited to, alkoxy, poly(ethylene glycol)-, alkyl-substituted amino, tetrahydrofuranyl, piperidinyl, morpholinyl, and the like.

[0052] Heteroaryl: The terms "heteroaryl" and "heteroar-", used alone or as part of a larger moiety, such as "heteroaralkyl" or "heteroaralkoxy", as used herein refer to a monocyclic, bicyclic, or polycyclic ring system having a total of 5 to 30 ring members, in which at least one ring in the system is aromatic and at least one aromatic ring atom is a heteroatom. In some embodiments, a heteroaryl group is a group having 5 to 10 ring atoms (i.e., monocyclic, bicyclic, or polycyclic), in some embodiments, 5, 6, 9, or 10 ring atoms. In some embodiments, each monocyclic ring unit is aromatic. In some embodiments, a heteroaryl group has 6, 10, or 14 pi electrons shared in a cyclic arrangement, and has 1 to 5 heteroatoms in addition to the carbon atoms. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. In some embodiments, a heteroaryl is a heterobiaryl group, such as bipyridyl. The terms "heteroaryl" and "heteroar-" as used herein also include groups in which a heteroaromatic ring is fused to one or more aryl, alicyclic, or heterocyclyl rings, with the group or point of attachment being on the heteroaromatic ring. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups can be monocyclic, bicyclic, or polycyclic.The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," any of which terms include rings that are optionally substituted. The term "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl group, where the alkyl and heteroaryl portions independently are optionally substituted.

[0053] Heteroatom: The term "heteroatom" as used herein means an atom that is not carbon or hydrogen. In some embodiments, the heteroatom is boron, oxygen, sulfur, nitrogen, phosphorus, or silicon (including oxidized forms of nitrogen, sulfur, phosphorus, or silicon; nitrogen (e.g., quaternized forms, forms such as iminium groups, etc.), phosphorus, sulfur, charged forms of oxygen, etc.). In some embodiments, the heteroatom is silicon, phosphorus, oxygen, sulfur, or nitrogen. In some embodiments, the heteroatom is silicon, oxygen, sulfur, or nitrogen. In some embodiments, the heteroatom is oxygen, sulfur, or nitrogen.

[0054] Heterocycle: As used herein, the terms "heterocycle", "heterocyclyl", "heterocyclic group", and "heterocyclic ring" are used interchangeably herein and refer to a monocyclic, bicyclic, or polycyclic ring moiety (e.g., 3-30 members) that is saturated or partially unsaturated and has one or more heteroatom ring atoms. In some embodiments, a heterocyclyl group is a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated and has, in addition to carbon atoms, one or more, preferably 1-4, heteroatoms as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur, and nitrogen, the nitrogen can be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or . +It may be NR (as in the case of N-substituted pyrrolidinyl). A heterocyclic ring may be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any of the ring atoms may be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle", "heterocyclyl", "heterocyclyl ring", "heterocyclic group", "heterocyclic moiety", and "heterocyclic group" are used interchangeably herein and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or alicyclic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl. Heterocyclyl groups can be monocyclic, bicyclic, or polycyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, where the alkyl and heterocyclyl portions are independently optionally substituted.

[0055] Identity: As used herein, the term "identity" refers to the overall relatedness between polymer molecules, e.g., between nucleic acid molecules (e.g., oligonucleotides, DNA, RNA, etc.) and / or between polypeptide molecules. In some embodiments, polymer molecules are considered to be "substantially identical" to each other if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical. Calculation of percent identity of two nucleic acid or polypeptide sequences can be performed, for example, by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced into one or both of the first and second sequences for optimal alignment, and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of the sequences aligned for comparison is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or substantially 100% of the length of the reference sequence. The nucleotides at corresponding positions are then compared. If a position in the first sequence is occupied by the same residue (e.g., nucleotide or amino acid) as the corresponding position in the second sequence, the molecules are identical at that position. The percent identity between two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps and the length of each gap that need to be introduced for optimal alignment of the two sequences. The sequence comparison and the determination of the percent identity between two sequences can be achieved using a mathematical algorithm. For example, the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller (CABIOS, 1989, 4:11-17) incorporated in the ALIGN program (version 2.0). In some exemplary embodiments, nucleic acid sequence comparisons performed with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4.Alternatively, the percent identity between two nucleotide sequences can be determined using the GAP program in the GCG software package using the NWSgapdna.CMP matrix.

[0056] Internucleotide bond: As used herein, the phrase "internucleotide bond" generally refers to a bond that links the nucleoside units of an oligonucleotide or nucleic acid. In some embodiments, the internucleotide bond is a phosphodiester bond (a natural phosphate bond (-OP(=O)(OH)O-), which may exist as a salt form, as will be understood by those of skill in the art) that is widely found in naturally occurring DNA and RNA molecules. In some embodiments, the internucleotide bond is a modified internucleotide bond (not a natural phosphate bond). In some embodiments, the internucleotide bond is a "modified internucleotide bond" in which at least one oxygen atom or -OH of the phosphodiester bond is replaced with a different organic or inorganic moiety. In some embodiments, such organic or inorganic moieties are =S, =Se, =NR', -SR', -SeR', -N(R'). 2 , B(R') 3, -S-, -Se-, and -N(R')-, where each R' is independently as defined and described in this disclosure. In some embodiments, the internucleotide linkage is a phosphotriester linkage, a phosphorothioate linkage (or a phosphorothioate diester linkage, -OP(=O)(SH)O-, which may exist as a salt form as understood by those skilled in the art) or a phosphorothioate triester linkage. In some embodiments, the modified internucleotide linkage is a phosphorothioate linkage. In some embodiments, the internucleotide linkage is, for example, one of a PNA (peptide nucleic acid) or a PMO (phosphorodiamidate morpholino oligomer) linkage. In some embodiments, the modified internucleotide linkage is a non-negatively charged internucleotide linkage. In some embodiments, the modified internucleotide linkage is a neutral internucleotide linkage (e.g., n001 in certain provided oligonucleotides). It will be understood by those skilled in the art that an internucleotide linkage may exist as an anion or cation at a given pH due to the presence of an acid or base moiety in the linkage. In some embodiments, the modified internucleotide linkages are the modified internucleotide linkages designated s, s1, s2, s3, s4, s5, s6, s7, s8, s9, s10, s11, s12, s13, s14, s15, s16, s17, and s18 as described in WO 2017 / 210647.

[0057] In vitro: As used herein, the term "in vitro" refers to events that take place not within a living organism (e.g., an animal, a plant, and / or a microorganism), but in an artificial environment, such as a test tube or reaction vessel, cell culture, etc.

[0058] In vivo: As used herein, the term "in vivo" refers to events that take place within an organism (e.g., an animal, a plant, and / or a microorganism).

[0059] Linked phosphorus: As defined herein, the phrase "linked phosphorus" is used to indicate that the particular phosphorus atom being referenced is a phosphorus atom present in an internucleotide linkage, which corresponds to the phosphorus atom of a phosphodiester internucleotide linkage present in naturally occurring DNA and RNA. In some embodiments, the linked phosphorus atom is present in a modified internucleotide linkage, in which each oxygen atom of the phosphodiester linkage is optionally and independently replaced by an organic or inorganic moiety. In some embodiments, the linked phosphorus atom is chiral (e.g., in the case of a phosphorothioate internucleotide linkage). In some embodiments, the linked phosphorus atom is achiral (e.g., as in a natural phosphate linkage).

[0060] Modified nucleobase: The term "modified nucleobase", "modified base" and the like refer to a chemical moiety that is chemically different from a nucleobase but can perform at least one function of a nucleobase. In some embodiments, a modified nucleobase is a nucleobase that includes a modification. In some embodiments, a modified nucleobase can perform at least one function of a nucleobase, for example, form a moiety in a polymer that can base pair with a nucleic acid that includes at least a complementary sequence of bases. In some embodiments, a modified nucleobase is a substituted A, T, C, G or U, or a substituted tautomer of A, T, C, G or U. In some embodiments, a modified nucleobase in the context of an oligonucleotide refers to a nucleobase that is not A, T, C, G or U.

[0061] Modified nucleosides: The term "modified nucleoside" refers to a moiety derived from or chemically similar to a natural nucleoside, but containing a chemical modification that distinguishes it from a natural nucleoside. Non-limiting examples of modified nucleosides include those that contain modifications at the base and / or sugar. Non-limiting examples of modified nucleosides include those that have a 2' modification at the sugar. Non-limiting examples of modified nucleosides also include abasic nucleosides (lacking a nucleobase). In some embodiments, modified nucleosides can perform at least one function of a nucleoside, such as forming a moiety in a polymer that can base pair with a nucleic acid that contains at least a complementary sequence of bases.

[0062] Modified Nucleotide: The term "modified nucleotide" includes any chemical moiety that is structurally different from a naturally occurring nucleotide, but can perform at least one function of a naturally occurring nucleotide. In some embodiments, a modified nucleotide comprises a modification at the sugar, base, and / or internucleotide linkage. In some embodiments, a modified nucleotide comprises a modified sugar, a modified nucleobase, and / or a modified internucleotide linkage. In some embodiments, a modified nucleotide can perform at least one function of a nucleotide, e.g., to form a subunit in a polymer that is capable of base pairing with a nucleic acid that comprises at least a complementary sequence of bases.

[0063] Modified sugar: The term "modified sugar" refers to a moiety that can replace a sugar. The modified sugar mimics the spatial arrangement, electronic properties, or some other physicochemical property of the sugar. In some embodiments, the modified sugar is a substituted ribose or deoxyribose, as described in this disclosure. In some embodiments, the modified sugar comprises a 2'-modification. Examples of useful 2'-modifications are widely available in the art and described herein. In some embodiments, the 2'-modification is 2'-F. In some embodiments, the 2'-modification is 2'-OR, where R is an optionally substituted C. 1~10In some embodiments, the 2'-modification is a 2'-OMe. In some embodiments, the 2'-modification is a 2'-MOE. In some embodiments, the modified sugar is a bicyclic sugar (e.g., a sugar used in LNA, BNA, etc.). In some embodiments, in the context of an oligonucleotide, the modified sugar is a sugar that is not a ribose or deoxyribose typically found in natural RNA or DNA.

[0064] Nucleic acid: The term "nucleic acid" as used herein includes any nucleotide and polymers thereof. The term "polynucleotide" as used herein refers to a polymeric form of nucleotides of any length, either ribonucleotides (RNA) or deoxyribonucleotides (DNA) or combinations thereof. These terms refer to the primary structure of the molecule and thus include double- and single-stranded DNA as well as double- and single-stranded RNA. These terms include, as equivalents, analogs of RNA or DNA, including modified nucleotides and / or modified polynucleotides, such as, but not limited to, through methylated, protected, and / or capped nucleotides or polynucleotides. These terms encompass poly- or oligo-ribonucleotides (RNA) and poly- or oligo-deoxyribonucleotides (DNA); RNA or DNA derived from nucleobases and / or N-glycosides or C-glycosides of modified nucleobases; nucleic acids derived from sugars and / or modified sugars; and nucleic acids derived from phosphate bridges and / or modified internucleotide linkages. The term encompasses nucleic acids containing any combination of nucleobases, modified nucleobases, sugars, modified sugars, phosphate bridges, or modified internucleotide linkages. Examples include, but are not limited to, nucleic acids containing a ribose moiety, nucleic acids containing deoxy-ribose, nucleic acids containing both a ribose moiety and a deoxyribose moiety, and nucleic acids containing a ribose moiety and a modified ribose moiety. Unless otherwise specified, the prefix poly- refers to a nucleic acid containing from 2 to about 10,000 nucleotide monomer units and the prefix oligo- refers to a nucleic acid containing from 2 to about 200 nucleotide monomer units.

[0065] Nucleobase: The term "nucleobase" refers to the portion of a nucleic acid that participates in hydrogen bonds that bind one nucleic acid strand to another complementary strand in a sequence-specific manner. The most common naturally occurring nucleobases are adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T). In some embodiments, naturally occurring nucleobases are modified adenine, guanine, uracil, cytosine, or thymine. In some embodiments, naturally occurring nucleobases are methylated adenine, guanine, uracil, cytosine, or thymine. In some embodiments, the nucleobase comprises a heteroaryl ring (where the ring atom is nitrogen) and, in a nucleoside, the nitrogen is attached to the sugar moiety. In some embodiments, the nucleobase comprises a heterocyclic ring (where the ring atom is nitrogen) and, in a nucleoside, the nitrogen is attached to the sugar moiety. In some embodiments, the nucleobase is a nucleobase other than the "modified nucleobases" adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T). In some embodiments, the modified nucleobase is a substituted A, T, C, G, or U. In some embodiments, the modified nucleobase is a substituted tautomer of A, T, C, G, or U. In some embodiments, the modified nucleobase is a methylated adenine, guanine, uracil, cytosine, or thymine. In some embodiments, the modified nucleobase mimics the spatial arrangement, electronic properties, or some other physicochemical properties of the nucleobase and retains the hydrogen bonding properties that bind one nucleic acid strand to another in a sequence-specific manner. In some embodiments, the modified nucleobase can pair with all five naturally occurring bases (uracil, thymine, adenine, cytosine, or guanine) without substantially affecting the melting behavior, recognition by intracellular enzymes or activities of the oligonucleotide duplex. As used herein, the term "nucleobase" also encompasses structural analogs that are used in place of natural or naturally occurring nucleotides, such as modified nucleobases and nucleobase analogs. In some embodiments, the nucleobase is an optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U.In some embodiments, "nucleobase" refers to a nucleobase unit in an oligonucleotide or nucleic acid (e.g., A, T, C, G, or U as in an oligonucleotide or nucleic acid).

[0066] Nucleoside: The term "nucleoside" refers to a moiety in which a nucleobase or modified nucleobase is covalently linked to a sugar or modified sugar. In some embodiments, the nucleoside is a natural nucleoside, such as adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, or deoxycytidine. In some embodiments, the nucleoside is a modified nucleoside, such as a substituted natural nucleoside selected from adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, and deoxycytidine. In some embodiments, the nucleoside is a substituted tautomer of a natural nucleoside selected from modified nucleosides, such as adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, and deoxycytidine. In some embodiments, "nucleoside" refers to a nucleoside unit in an oligonucleotide or nucleic acid.

[0067] Nucleotide: The term "nucleotide" as used herein refers to a monomeric unit of a polynucleotide consisting of a nucleobase, a sugar, and one or more internucleotide linkages (e.g., phosphate linkages in natural DNA and RNA). Naturally occurring bases [guanine, (G), adenine, (A), cytosine, (C), thymine, (T), and uracil (U)] are derivatives of purines or pyrimidines, but should be understood to include naturally occurring and non-naturally occurring base analogs. Naturally occurring sugars are the pentose (five-carbon sugars) deoxyribose (forming DNA) or ribose (forming RNA), but should be understood to include naturally occurring and non-naturally occurring sugar analogs. Nucleotides are linked via internucleotide linkages to form nucleic acids or polynucleotides. Many internucleotide linkages are known in the art (e.g., but are not limited to, phosphate, phosphorothioate, boranophosphate, etc.). Artificial nucleic acids include PNA (peptide nucleic acid), phosphotriester, phosphorothioate, H-phosphonate, phosphoramidate, boranophosphate, methylphosphonate, phosphonoacetate, thiophosphonoacetate, and other variants of the phosphate backbone of natural nucleic acids, such as those described herein. In some embodiments, natural nucleotides include naturally occurring bases, sugars, and internucleotide bonds. As used herein, the term "nucleotide" also encompasses structural analogs that are used in place of natural or naturally occurring nucleotides, such as modified nucleotides and nucleotide analogs. In some embodiments, "nucleotide" refers to a nucleotide unit in an oligonucleotide or nucleic acid.

[0068] Oligonucleotide: The term "oligonucleotide" refers to a polymer or oligomer of nucleotides, which may contain any combination of natural and unnatural nucleobases, sugars, and internucleotide linkages.

[0069] Oligonucleotides can be single-stranded or double-stranded. Single-stranded oligonucleotides can have a double-stranded region (formed by two parts of single-stranded oligonucleotides), and double-stranded oligonucleotides that contain two oligonucleotide strands can have a single-stranded region, for example, in the region where the two oligonucleotide strands are not complementary to each other. Examples of oligonucleotides include, but are not limited to, structural genes, genes including control and termination regions, self-replicating systems such as viruses or plasmid DNA, single-stranded and double-stranded RNAi agents and other RNA interference reagents (RNAi agents or iRNA agents), shRNA, antisense oligonucleotides, ribozymes, microRNA, microRNA mimics, supermir, aptamers, antimir, antagomir, Ul adaptors, triplex-forming oligonucleotides, G-quadruplex oligonucleotides, RNA activators, immunostimulatory oligonucleotides, and decoy oligonucleotides.

[0070] The oligonucleotides of the present disclosure can be of various lengths. In certain embodiments, the oligonucleotides can range from about 2 to about 200 nucleosides in length. In various related embodiments, the single-stranded, double-stranded, or triple-stranded oligonucleotides can range in length from about 4 to about 10 nucleosides, about 10 to about 50 nucleosides, about 20 to about 50 nucleosides, about 15 to about 30 nucleosides, about 20 to about 30 nucleosides in length. In some embodiments, the oligonucleotides are about 9 to about 39 nucleosides in length. In some embodiments, the oligonucleotides are about 25 to about 70 nucleosides in length. In some embodiments, the oligonucleotides are about 26 to about 70 nucleosides in length. In some embodiments, the oligonucleotides are about 27 to about 70 nucleosides in length. In some embodiments, the oligonucleotides are about 28 to about 70 nucleosides in length. In some embodiments, the oligonucleotides are about 29 to about 70 nucleosides in length. In some embodiments, the oligonucleotide is about 30 to about 70 nucleosides in length. In some embodiments, the oligonucleotide is about 31 to about 70 nucleosides in length. In some embodiments, the oligonucleotide is about 32 to about 70 nucleosides in length. In some embodiments, the oligonucleotide is about 25 to about 60 nucleosides in length. In some embodiments, the oligonucleotide is about 25 to about 50 nucleosides in length. In some embodiments, the oligonucleotide is about 25 to about 40 nucleosides in length. In some embodiments, the oligonucleotide is about 30 to about 40 nucleosides in length. In some embodiments, the oligonucleotide is at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleosides in length. In some embodiments, the oligonucleotide is at least 4 nucleosides in length. In some embodiments, the oligonucleotide is at least 5 nucleosides in length. In some embodiments, the oligonucleotide is at least 6 nucleosides in length. In some embodiments, the oligonucleotide is at least 7 nucleosides in length.In some embodiments, the oligonucleotide is at least 8 nucleosides in length. In some embodiments, the oligonucleotide is at least 9 nucleosides in length. In some embodiments, the oligonucleotide is at least 10 nucleosides in length. In some embodiments, the oligonucleotide is at least 11 nucleosides in length. In some embodiments, the oligonucleotide is at least 12 nucleosides in length. In some embodiments, the oligonucleotide is at least 15 nucleosides in length. In some embodiments, the oligonucleotide is at least 15 nucleosides in length. In some embodiments, the oligonucleotide is at least 16 nucleosides in length. In some embodiments, the oligonucleotide is at least 17 nucleosides in length. In some embodiments, the oligonucleotide is at least 18 nucleosides in length. In some embodiments, the oligonucleotide is at least 19 nucleosides in length. In some embodiments, the oligonucleotide is at least 20 nucleosides in length. In some embodiments, the oligonucleotide is at least 25 nucleosides in length. In some embodiments, the oligonucleotide is at least 26 nucleosides in length. In some embodiments, the oligonucleotide is at least 27 nucleosides in length. In some embodiments, the oligonucleotide is at least 28 nucleosides in length. In some embodiments, the oligonucleotide is at least 29 nucleosides in length. In some embodiments, the oligonucleotide is at least 30 nucleosides in length. In some embodiments, the oligonucleotide is at least 31 nucleosides in length. In some embodiments, the oligonucleotide is at least 32 nucleosides in length. In some embodiments, the oligonucleotide is at least 33 nucleosides in length. In some embodiments, the oligonucleotide is at least 34 nucleosides in length. In some embodiments, the oligonucleotide is at least 35 nucleosides in length. In some embodiments, the oligonucleotide is at least 36 nucleosides in length.In some embodiments, the oligonucleotide is at least 37 nucleosides in length. In some embodiments, the oligonucleotide is at least 38 nucleosides in length. In some embodiments, the oligonucleotide is at least 39 nucleosides in length. In some embodiments, the oligonucleotide is at least 40 nucleosides in length. In some embodiments, the oligonucleotide is 25 nucleosides in length. In some embodiments, the oligonucleotide is 26 nucleosides in length. In some embodiments, the oligonucleotide is 27 nucleosides in length. In some embodiments, the oligonucleotide is 28 nucleosides in length. In some embodiments, the oligonucleotide is 29 nucleosides in length. In some embodiments, the oligonucleotide is 30 nucleosides in length. In some embodiments, the oligonucleotide is 31 nucleosides in length. In some embodiments, the oligonucleotide is 32 nucleosides in length. In some embodiments, the oligonucleotide is 33 nucleosides in length. In some embodiments, the oligonucleotide is 34 nucleosides in length. In some embodiments, the oligonucleotide is 35 nucleosides in length. In some embodiments, the oligonucleotide is 36 nucleosides in length. In some embodiments, the oligonucleotide is 37 nucleosides in length. In some embodiments, the oligonucleotide is 38 nucleosides in length. In some embodiments, the oligonucleotide is 39 nucleosides in length. In some embodiments, the oligonucleotide is 40 nucleosides in length. In some embodiments, each nucleoside counted in the oligonucleotide length independently comprises a nucleobase that comprises a ring having at least one nitrogen ring atom. In some embodiments, each nucleoside counted in the oligonucleotide length independently comprises A, T, C, G, or U, or an optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U.

[0071] Oligonucleotide type: As used herein, the phrase "oligonucleotide type" is used to define oligonucleotides having a particular base sequence, backbone linkage pattern (i.e., pattern of internucleotide linkage types, e.g., phosphate, phosphorothioate, phosphorothioate triester, etc.), pattern of backbone chiral centers [i.e., pattern of phosphorus stereochemistry (Rp / Sp)], and pattern of backbone phosphorus modifications. In some embodiments, oligonucleotides of a commonly named "type" are structurally identical to one another.

[0072] Those skilled in the art will appreciate that the synthesis methods of the present disclosure provide a degree of control during the synthesis of an oligonucleotide chain, such that each nucleotide unit of the oligonucleotide chain can be designed and / or selected in advance to have a particular stereochemistry at the binding phosphorus and / or a particular modification at the binding phosphorus, and / or a particular base, and / or a particular sugar. In some embodiments, the oligonucleotide chain is designed and / or selected in advance to have a particular combination of stereocenters at the binding phosphorus. In some embodiments, the oligonucleotide chain is designed and / or determined to have a particular combination of modifications at the binding phosphorus. In some embodiments, the oligonucleotide chain is designed and / or selected to have a particular combination of bases. In some embodiments, the oligonucleotide chain is designed and / or selected to have a particular combination of one or more of the structural features described above. In some embodiments, the present disclosure provides compositions (e.g., chiral controlled oligonucleotide compositions) that include or consist of a plurality of oligonucleotide molecules. In some embodiments, all such molecules are of the same type (i.e., structurally identical to one another). However, in some embodiments, the compositions provided typically include a plurality of oligonucleotides of different types in predetermined relative amounts.

[0073] Optionally substituted: As described herein, the compounds of the present disclosure, such as oligonucleotides, may contain optionally substituted and / or substituted moieties. In general, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogens of the specified moiety are replaced with a suitable substituent. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at all positions. In some embodiments, an optionally substituted group is unsubstituted. The combinations of substituents envisioned by the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds. As used herein, the term "stable" refers to compounds that are substantially unchanged when subjected to conditions that allow for their production, detection, and, in certain embodiments, their harvesting, purification, and use for one or more of the purposes disclosed herein. Particular substituents are described below.

[0074] Suitable monovalent substituents on substitutable atoms, e.g., suitable carbon atoms, are independently halogen; -(CH 2 ) 0~4 R°;-(CH 2 ) 0~4 OR°;-O(CH 2 ) 0~4 R°, -O-(CH 2 ) 0~4 C(O)OR°;-(CH 2 ) 0~4 CH(OR°) 2 ;-(CH 2 ) 0~4 Ph (optionally substituted with R°); -(CH 2 ) 0~4 O(CH 2 ) 0~1 Ph (optionally substituted with R°); -CH=CHPh (optionally substituted with R°); -(CH 2 ) 0~4 O(CH 2 )0~1 -pyridyl (which may be substituted by R°); -NO 2 ; -CN; -N 3 ; -(CH 2 ) 0~4 N(R°) 2 ; -(CH 2 ) 0~4 N(R°)C(O)R°; -N(R°)C(S)R°; -(CH 2 ) 0~4 N(R°)C(O)NR° 2 ; -N(R°)C(S)NR° 2 ; -(CH 2 ) 0~4 N(R°)C(O)OR°; -N(R°)N(R°)C(O)R°; -N(R°)N(R°)C(O)NR° 2 ; -N(R°)N(R°)C(O)OR°; -(CH 2 ) 0~4 C(O)R°; -C(S)R°; -(CH 2 ) 0~4 C(O)OR°; -(CH 2 ) 0~4 C(O)SR°; -(CH 2 ) 0~4 C(O)OSiR° 3 ; -(CH 2 ) 0~4 OC(O)R°; -OC(O)(CH 2 ) 0~4 SR°, -SC(S)SR°; -(CH 2 ) 0~4 SC(O)R°; -(CH 2 ) 0~4 C(O)NR° 2 ; -C(S)NR° 2 ; -C(S)SR°; -(CH 2 ) 0~4 OC(O)NR° 2 ; -C(O)N(OR°)R°; -C(O)C(O)R°; -C(O)CH 2 C(O)R°; -C(NOR°)R°; -(CH 2 ) 0-4 SSR°; -(CH 2 ) 0~4 S(O) 2 R°; -(CH 2 ) 0~4 S(O)2 OR°;-(CH 2 ) 0~4 OS(O) 2 R°;-S(O) 2 NR° 2 ;-(CH 2 ) 0-4 S(O)R°;-N(R°)S(O) 2 NR° 2 ;-N(R°)S(O) 2 R°;-N(OR°)R°;-C(NH)NR° 2 ;-Si(R°) 3 ;-OSi(R°) 3 ;-B(R°) 2 ;-OB(R°) 2 ;-OB(OR°) 2 ;-P(R°) 2 ;-P(OR°) 2 ;-P(R°)(OR°);-OP(R°) 2 ;-OP(OR°) 2 ;-OP(R°)(OR°);-P(O)(R°) 2 ;-P(O)(OR°) 2 ;-OP(O)(R°) 2 ;-OP(O)(OR°) 2 ;-OP(O)(OR°)(SR°);-SP(O)(R°) 2 ;-SP(O)(OR°) 2 ;-N(R°)P(O)(R°) 2 ;-N(R°)P(O)(OR°) 2 ;-P(R°) 2 [B(R°) 3 ];-P(OR°) 2 [B(R°) 3 ];-OP(R°) 2 [B(R°) 3 ];-OP(OR°) 2 [B(R°) 3 ];-(C 1~4 Linear or branched alkylene) ON (R°) 2 ;or-(C 1~4 Linear or branched alkylene)C(O)ON(R°) 2 wherein each R° may be optionally substituted as defined herein and is independently hydrogen, C 1~20C having 1 to 5 heteroatoms independently selected from aliphatic, nitrogen, oxygen, sulfur, silicon and phosphorus 1~20 Heteroaliphatic, -CH 2 -(C 6~14 Aryl), -O(CH 2 ) 0~1 (C 6~14 Aryl), -CH 2 -(5-14 membered heteroaryl ring), a 5-20 membered monocyclic, bicyclic or polycyclic saturated, partially unsaturated or aryl ring having 0-5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon and phosphorus, or, notwithstanding the above definition, two independent occurrences of R° together with their intervening atoms form a 5-20 membered monocyclic, bicyclic or polycyclic saturated, partially unsaturated or aryl ring (which may be substituted as defined below) having 0-5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon and phosphorus.

[0075] Suitable monovalent substituents on R° (or the ring formed by two independent occurrences of R° together with their intervening atoms) are independently halogen, -(CH 2 ) 0~2 R ● , -(Halo R ● ), -(CH 2 ) 0~2 OH, -(CH 2 ) 0~2 OR ● , -(CH 2 ) 0~2 CH(OR ● ) 2 ;-O(Halo R ● ), -CN, -N 3 , -(CH 2 ) 0~2 C(O)R ● , -(CH 2 ) 0~2 C(O)OH, -(CH 2 ) 0~2 C(O)OR ● , -(CH 2 ) 0~2 S.R. ● , -(CH 2 ) 0~2 SH, -(CH2 ) 0~2 NH 2 , -(CH 2 ) 0~2 NHR ● , -(CH 2 ) 0~2 NR ● 2 , -NO 2 , -SiR ● 3 , -OSiR ● 3 , -C(O)SR ● , -(C 1~4 Linear or branched alkylene)C(O)OR ● , or -SSR ● where each R ● is unsubstituted or, if preceded by "halo", is substituted only with one or more halogens, and C 1~4 Aliphatic, -CH 2 Ph, -O(CH 2 ) 0~1 and a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. Suitable divalent substituents on a saturated carbon atom of R include =O and =S.

[0076] For example, suitable divalent substituents on suitable carbon atoms are, independently, the following: =O, =S, =NNR * 2 , =NNHC(O)R * , =NNHC(O)OR * , =NNHS(O) 2 R * , =NR * , =NOR * , -O(C(R * 2 )) 2~3 O- or -S(C(R * 2 )) 2~3 S-, where R * Each independent occurrence of is hydrogen, C which may be substituted as defined below. 1~6Aliphatic and unsubstituted 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur. Suitable divalent substituents attached to adjacent substitutable carbons of an "optionally substituted" group include -O(CR * 2 ) 2~3 O-, where R * Each independent occurrence of is hydrogen, C which may be substituted as defined below. 1~6 It is selected from aliphatic and unsubstituted 5-6 membered saturated rings, partially unsaturated rings, and aryl rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0077] R * Suitable substituents on the aliphatic groups are independently halogen, -R ● , -(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH 2 , -NHR ● , -NR ● 2 OR -NO 2 where each R ● is unsubstituted or, when preceded by "halo", is substituted with one or more halogens only, and independently represents C 1~4 Aliphatic, -CH 2 Ph, -O(CH 2 ) 0~1 Ph, or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0078] In some embodiments, suitable substituents on a substitutable nitrogen are independently -R † , -NR † 2 , -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CH 2 C(O)R† , -S(O) 2 R † , -S(O) 2 NR † 2 , -C(S)NR † 2 , -C(NH)NR † 2 Or -N(R † )S(O) 2 R † where each R † are independently hydrogen, C which may be substituted as defined below 1~6 an aliphatic, unsubstituted -OPh, or an unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or, notwithstanding the above definitions, R † two independent occurrences of, taken together with their intervening atoms, form an unsubstituted 3-12 membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur.

[0079] R † Suitable substituents on the aliphatic groups are independently halogen, -R ● , -(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH 2 , -NHR ● , -NR ● 2 OR -NO 2 where each R ● is unsubstituted or, when preceded by "halo", is substituted with one or more halogens only, and independently represents C 1~4 Aliphatic, -CH 2 Ph, -O(CH 2 ) 0~1 Ph, or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0080] P-modification: As used herein, the term "P-modification" refers to any modification at the bound phosphorus other than a stereochemical modification. In some embodiments, a P-modification includes the addition, substitution, or removal of a pendant moiety covalently attached to the bound phosphorus.

[0081] Partially unsaturated: As used herein, the term "partially unsaturated" refers to a ring moiety that contains at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings with multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as defined herein.

[0082] Pharmaceutical composition: As used herein, the term "pharmaceutical composition" refers to an active agent formulated with one or more pharma-ceutically acceptable carriers. In some embodiments, the active agent is present in a unit dose suitable for administration in a treatment regimen that exhibits a statistically significant probability of achieving a predetermined therapeutic effect when administered to an appropriate population. In some embodiments, the pharmaceutical composition can be specially formulated for administration in solid or liquid form, including those compatible with the following: oral administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., buccal, sublingual and those targeting systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, e.g., as a sterile solution or suspension or sustained release formulation, e.g., by subcutaneous, intramuscular, intravenous or epidural injection; topical application, e.g., as a cream, ointment or controlled release patch or spray applied to the skin, lungs or oral cavity; vaginal or rectal, e.g., as a suppository, cream or foam; sublingual; ocular; transdermal; or intranasal, pulmonary and other mucosal surfaces.

[0083] Pharmaceutically acceptable: As used herein, the phrase "pharmacologically acceptable" refers to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0084] Pharmaceutically acceptable carrier: As used herein, the term "pharmaceutical acceptable carrier" means a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent that encapsulates a material, which is involved in carrying or transporting a compound of interest from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which may serve as pharma- ceutically acceptable carriers include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; pH buffer solutions; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic, compatible substances utilized in pharmaceutical formulations.

[0085] Pharmaceutically acceptable salts: The term "pharmaceutical acceptable salts" as used herein refers to salts of such compounds suitable for use in a pharmaceutical context, i.e., salts suitable for use in contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., within the scope of sound medical judgment, and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al. provide a detailed description of pharmaceutical acceptable salts in J. Pharmaceutical Sciences, 66:1-19 (1977). In some embodiments, pharmaceutical acceptable salts include, but are not limited to, non-toxic acid addition salts that are salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods used in the art, such as ion exchange. In some embodiments, pharma- ceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, and 2-hydroxy-ethanesulfonate. , lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like.In some embodiments, provided compounds contain one or more acidic groups (e.g., oligonucleotides), and the pharma- ceutically acceptable salt is an alkali metal salt, an alkaline earth metal salt, or an ammonium salt (e.g., N(R). 3where each R is independently defined and described in this disclosure). Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. In some embodiments, the pharma- ceutically acceptable salt is a sodium salt. In some embodiments, the pharma- ceutically acceptable salt is a potassium salt. In some embodiments, the pharma- ceutically acceptable salt is a calcium salt. In some embodiments, the pharma- ceutical acceptable salt includes non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyls having 1-6 carbon atoms, sulfonates, and arylsulfonates, as appropriate. In some embodiments, provided compounds include two or more acid groups, e.g., oligonucleotides may include two or more acid groups (e.g., in natural phosphate linkages and / or modified internucleotide linkages). In some embodiments, pharma- ceutical acceptable salts, or salts in general, of such compounds include two or more cations, which may be the same or different. In some embodiments, in a pharma- ceutically acceptable salt (or salt in general), all ionizable hydrogens in acidic groups (e.g., in an aqueous solution having a pKa of about 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2 or less, in some embodiments, about 7 or less; in some embodiments, about 6 or less; in some embodiments, about 5 or less; in some embodiments, about 4 or less; in some embodiments, about 3 or less) are replaced with cations. In some embodiments, each phosphorothioate and phosphate group is independently present in its salt form (e.g., in the case of a sodium salt, -OP(O)(SNa)-O- and -OP(O)(ONa)-O-, respectively). In some embodiments, each phosphorothioate and phosphate internucleotide linkage is independently present in its salt form (e.g., in the case of a sodium salt, -OP(O)(SNa)-O- and -OP(O)(ONa)-O-, respectively). In some embodiments, the pharma- ceutically acceptable salt is a sodium salt of an oligonucleotide.In some embodiments, the pharma- ceutically acceptable salt is a sodium salt of the oligonucleotide, wherein each acidic phosphate group and modified phosphate group (e.g., phosphorothioate, phosphate, etc.), if present, is present as a salt form (all sodium salts).

[0086] Predetermined: Predetermined (or pre-determined) means, for example, selected or non-random or controlled, as opposed to randomly occurring, random, or achieved without control. Those skilled in the art reading this specification will understand that the present disclosure provides techniques that allow for the selection of specific chemical and / or stereochemical features to be incorporated into an oligonucleotide composition, and further allows for the controlled preparation of oligonucleotide compositions having such chemical and / or stereochemical features. A composition so provided is "predetermined" as described herein. A composition that may contain a particular oligonucleotide is not a "predetermined" composition because it was accidentally generated through a process that is not controlled to intentionally generate a specific chemical and / or stereochemical feature. In some embodiments, a predetermined composition is one that can be reproduced intentionally (e.g., by repetition of a controlled process). In some embodiments, a predetermined level of a plurality of oligonucleotides in a composition means that the absolute amount and / or the relative amount (ratio, percentage, etc.) of the plurality of oligonucleotides in the composition is controlled. In some embodiments, a predetermined level of a plurality of oligonucleotides in a composition is achieved by the preparation of chiral controlled oligonucleotides.

[0087] Protecting Group: The term "protecting group" as used herein is a group that is well known in the art and is described in Organic Synthesis, TW Greene and PGM Wets, 3, incorporated herein by reference in its entirety. rdProtective Groups include those described in detail in Protecting Groups, New York: The American Chemical Society, edition, John Wiley & Sons, 1999. Also included are those protecting groups specifically adapted for nucleoside and nucleotide chemistry described in Current Protocols in Nucleic Acid Chemistry 06 / 2012, edited by Serge L. Beaucage et al., Chapter 2 of which is incorporated herein by reference in its entirety. Suitable amino-protecting groups include methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxantyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate (Teoc). , 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc), 1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2'- and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-Dicyclohexylcarboxamido)ethyl, t-Butyl carbamate (BOC), 1-Adamantyl carbamate (Adoc), Vinyl carbamate (Voc), Allyl carbamate (Alloc), 1-Isopropylallyl carbamate (Ipaoc), Cinnamyl carbamate (Coc), 4-Nitrocinnamyl carbamate (Noc), 8-Quinolyl carbamate, N-Hydroxypiperidinyl carbamate, Carbamate Carbamic acid alkyldithio, carbamic acid benzyl (Cbz), carbamic acid p-methoxybenzyl (Moz), carbamic acid p-nitrile (nitobenzyl), carbamic acid p-bromobenzyl, carbamic acid p-chlorobenzyl, carbamic acid 2,4-dichlorobenzyl, carbamic acid 4-methylsulfinylbenzyl (Msz), carbamic acid 9-anthrylmethyl, carbamic acid diphenylmethyl, carbamic acid 2 -Methylthioethyl, 2-methylsulfonylethyl carbamate, 2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methyl carbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3,4-Dimethoxy-6-nitrobenzyl, o-nitrophenyl phenyl carbamate, phenothiazinyl-(10)-carbonyl derivatives, N'-p-toluenesulfonylaminocarbonyl derivatives, N'-phenylaminothiocarbonyl derivatives, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate , 2,2-dimethoxycarbonylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate, 1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate, 2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynnyl carbamate, isobutyl carbamate, isonicotinyl carbamate, p-(p'-methoxyphenyl) carbamate p-phenylazo)benzyl, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-t- Butylphenyl, 4-(trimethylammonium)benzyl carbamate, 2,4,6-trimethylbenzyl carbamate, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide,(N'-dithiobenzyloxycarbonylamino)acetamide, 3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinamide, N-acetylmethionine derivatives, o-nitrobenzamide, o-(benzoyloxymethyl)benzamide, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3 -Acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N -2-Picolylamino N'-oxide, N-1,1-dimethylthiomethyleneamine, N-benzylideneamine, Np-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine, N-(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine, Np-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylideneamine, N-(5,5-Dimethyl-3-oxo-1-cyclohexenyl)amine, N-borane derivatives, N-diphenylborinic acid derivatives, N-[phenyl(pentacarbonylchromium- or tungsten)carbonyl]amine, N-copper chelates, N-zinc chelates, N-nitroamines, N-nitrosamines, amine N-oxides, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidate, dibenzyl phosphoramidate, diphenyl phosphoramidate, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, 3-nitropyridine sulfenamide (Npys), p-toluenesulfonamide (Ts), benzene Sulfonamides, 2,3,6-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4 -methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

[0088] Suitable protected carboxylic acids further include, but are not limited to, silyl-, alkyl-, alkenyl-, aryl-, and arylalkyl-protected carboxylic acids. Examples of suitable silyl groups include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl, and the like. Examples of suitable alkyl groups include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, tetrahydropyran-2-yl. Examples of suitable alkenyl groups include allyl. Examples of suitable aryl groups include optionally substituted phenyl, biphenyl, or naphthyl. Examples of suitable arylalkyl groups include optionally substituted benzyl (e.g., p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl), and 2- and 4-picolyl.

[0089] Suitable hydroxyl protecting groups include methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (M EM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl] -4-Methoxypiperidin-4-yl (CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trimethyl-1-phenylpropanediol, 1- ... ethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxide, diphenylmethyl, p,p'-Dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromophenacyloxyphenyl)diphenylmethyl, 4,4',4''-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4',4''-tris(levulinoyloxyphenyl)methyl, 4,4',4''-tris(benzoyl) oxyphenyl)methyl, 3-(imidazol-1-yl)bis(4',4''-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodithiolan-2-yl, benzisothiazolyl S,S-dioxide, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethysilyl Diacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate, adamanoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2,2,2-Trichloroethyl (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphonio)ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate, alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate, 4-ethoxy-1-naphthoyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methyl (ethylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinate, (E)-2-methyl-2-butenoate, o-(methoxycarbonyl)benzoate, α-naphthoate, nitrate, alkyl N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts). To protect 1,2- or 1,3-diols, the following protecting groups may be used: methylene acetal, ethylidene acetal, 1-t-butylethylidene ketal, 1-phenylethylidene ketal, (4-methoxyphenyl)ethylidene acetal, 2,2,2-trichloroethylidene acetal, acetonide, cyclopentylidene ketal, cyclohexylidene ketal, cycloheptylidene ketal, benzylidene acetal, p-methoxybenzylidene acetal, 2,4-dimethoxybenzylidene ketal, 3,4-dimethoxybenzylidene acetal, 2-nitrobenzylidene acetal, methoxymethylene acetal, ethoxymethylene acetal, dimethoxymethylene orthoester, 1-methoxyethylidene orthoester, 1-ethoxyethylidene orthoester, 1,2-dimethoxyethylidene orthoester, α-methoxybenzylidene orthoester, 1-(N,N-dimethylamino)ethylidene derivative, α-(N,N'-dimethylamino)benzylidene derivative, 2-oxacyclopentylidene orthoester, di-t-butylsilylene group (DTBS), 1,3-(1,1,3,3-tetraisopropyldisiloxanylidene) derivative (TIPDS), tetra-t-butoxydisiloxane-1,3-diylidene derivative (TBDS), cyclic carbonate, cyclic boronate, ethyl borate, and phenyl borate.

[0090] In some embodiments, the hydroxyl protecting group is acetyl, t-butyl, t-butoxymethyl, methoxymethyl, tetrahydropyranyl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 2-trimethylsilylethyl, p-chlorophenyl, 2,4-dinitrophenyl, benzyl, benzoyl, p-phenylbenzoyl, 2,6-dichlorobenzyl, diphenylmethyl, p-nitrobenzyl, triphenylmethyl(trityl), 4,4'-dimethoxytrityl, trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triphenylsilyl, triisopropylsilyl, benzoylformate, chloroacetyl, trichloroacetyl, trifiuoroacetyl, pivaloyl, 9-fluorenylmethyl carbonate, mesylate, tosylate, triflate, trityl, monomethoxytrityl (MMTr), 4,4'-dimethoxytrityl, xyltrityl, (DMTr) and 4,4',4''-trimethoxytrityl (TMTr), 2-cyanoethyl (CE or Cne), 2-(trimethylsilyl)ethyl (TSE), 2-(2-nitrophenyl)ethyl, 2-(4-cyanophenyl)ethyl 2-(4-nitrophenyl)ethyl (NPE), 2-(4-nitrophenylsulfonyl)ethyl, 3,5-dichlorophenyl, 2,4-dimethylphenyl, 2-nitrophenyl, 4-nitrophenyl , 2,4,6-trimethylphenyl, 2-(2-nitrophenyl)ethyl, butylthiocarbonyl, 4,4',4''-tris(benzoyloxy)trityl, diphenylcarbamoyl, levulinyl, 2-(dibromomethyl)benzoyl (Dbmb), 2-(isopropylthiomethoxymethyl)benzoyl (Ptmt), 9-phenylxanthen-9-yl (Pixyl), or 9-(p-methoxyphenyl)xanthin-9-yl (MOX). In some embodiments, each of the hydroxyl protecting groups is independently selected from acetyl, benzyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, and 4,4'-dimethoxytrityl. In some embodiments, the hydroxyl protecting group is selected from the group consisting of trityl, monomethoxytrityl, and 4,4'-dimethoxytrityl groups.In some embodiments, the phosphorus-linked protecting group is a group that is attached to a phosphorus linkage (e.g., an internucleotide linkage) throughout oligonucleotide synthesis. In some embodiments, the protecting group is attached to the sulfur atom of a phosphorothioate group. In some embodiments, the protecting group is attached to the oxygen atom of an internucleotide phosphorothioate linkage. In some embodiments, the protecting group is attached to the oxygen atom of an internucleotide phosphate linkage. In some embodiments, the protecting group is 2-cyanoethyl (CE or Cne), 2-trimethylsilylethyl, 2-nitroethyl, 2-sulfonylethyl, methyl, benzyl, o-nitrobenzyl, 2-(p-nitrophenyl)ethyl (NPE or Npe), 2-phenylethyl, 3-(N-tert-butylcarboxamido)-1-propyl, 4-oxopentyl, 4-methylthio-1-butyl, 2-cyano-1,1-dimethylethyl, 4-N-methylaminobutyl, 3-(2-pyridyl)-1-propyl, 2-[N-methyl-N-(2-pyridyl)]aminoethyl, 2-(N-formyl,N-methyl)aminoethyl, or 4-[N-methyl-N-(2,2,2-trifluoroacetyl)amino]butyl.

[0091] Subject: As used herein, the term "subject" or "test subject" refers to any organism to which a compound (e.g., an oligonucleotide) or composition is administered, e.g., for experimental, diagnostic, prophylactic, and / or therapeutic purposes, in accordance with the present disclosure. Exemplary subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans; insects; parasites, etc.), and plants. In some embodiments, the subject is a human. In some embodiments, the subject may be suffering from and / or susceptible to a disease, disorder, and / or condition.

[0092] Substantially: As used herein, the term "substantially" refers to the qualitative condition of exhibiting the entire or nearly entire extent or degree of a property or characteristic of interest. A base sequence that is substantially identical or complementary to a second sequence is not completely identical or complementary to the second sequence, but is largely or nearly identical or complementary to the second sequence. In some embodiments, an oligonucleotide having a sequence that is substantially complementary to another oligonucleotide or nucleic acid forms a duplex with the oligonucleotide or nucleic acid in a manner similar to an oligonucleotide having a completely complementary sequence. In addition, those skilled in the art of biology and / or chemistry will understand that biological and chemical phenomena rarely, if ever, proceed to completion and / or perfection or achieve or avoid absolute results. Thus, the term "substantially" as used herein is used to capture the potential lack of completeness inherent in many biological and / or chemical phenomena.

[0093] Sugar: The term "sugar" refers to closed and / or open monosaccharides or polysaccharides. In some embodiments, the sugar is a monosaccharide. In some embodiments, the sugar is a polysaccharide. Sugars include, but are not limited to, ribose, deoxyribose, pentofuranose, pentopyranose, and hexopyranose moieties. As used herein, the term "sugar" also encompasses structural analogs that are used in place of traditional sugar molecules, such as glycols, polymers that form the backbone of nucleic acid analogs, glycol nucleic acids ("GNAs"). As used herein, the term "sugar" also encompasses structural analogs that are used in place of natural or naturally occurring nucleotides, such as modified sugars and nucleotide sugars. In some embodiments, the sugar is an RNA or DNA sugar (ribose or deoxyribose). In some embodiments, the sugar is a modified ribose or deoxyribose sugar (e.g., 2'-modified, 5'-modified, etc.). As described herein, in some embodiments, when used in oligonucleotides and / or nucleic acids, modified sugars can provide one or more desired properties, activities, etc. In some embodiments, the sugar is an optionally substituted ribose or deoxyribose. In some embodiments, "sugar" refers to a sugar unit in an oligonucleotide or nucleic acid.

[0094] Susceptible: An individual who is "susceptible" to a disease, disorder, and / or condition is an individual who is at a higher risk of developing the disease, disorder, and / or condition than the general population. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition has a predisposition to having the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may not have been diagnosed with the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may exhibit symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may not exhibit symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition will develop the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition will not develop the disease, disorder, and / or condition.

[0095] Therapeutic Agent: As used herein, the term "therapeutic agent" generally refers to any agent that induces a desired effect (e.g., a desired biological, clinical, or pharmacological effect) when administered to a subject. In some embodiments, an agent is considered to be a therapeutic agent if it exhibits a statistically significant effect across a suitable population. In some embodiments, the suitable population is a population of subjects suffering from and / or susceptible to a disease, disorder, or condition. In some embodiments, the suitable population is a population of model organisms. In some embodiments, the suitable population may be defined by one or more criteria, such as age group, sex, genetic background, pre-existing clinical conditions, etc., prior to receiving therapy. In some embodiments, a therapeutic agent is a substance that, when administered to a subject in an effective amount, relieves, ameliorates, alleviates, inhibits, prevents, delays the onset of, reduces the severity of, and / or reduces the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition of the subject. In some embodiments, a "therapeutic agent" is an agent that has been approved or is required to be approved by a government agency before it can be marketed for administration to humans. In some embodiments, a "therapeutic agent" is a drug for which a prescription is required for administration to a human. In some embodiments, a therapeutic agent is a provided compound, such as a provided oligonucleotide.

[0096] Therapeutically effective amount: As used herein, the term "therapeutically effective amount" refers to an amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that induces a desired biological response when administered as part of a treatment regimen. In some embodiments, a therapeutically effective amount of a substance is an amount sufficient to treat, diagnose, prevent, and / or delay the onset of a disease, disorder, and / or condition when administered to a subject suffering from or susceptible to the disease, disorder, and / or condition. As will be appreciated by one of skill in the art, the effective amount of a substance can vary depending on factors such as the desired biological endpoint, the substance delivered, the target cell or tissue, and the like. For example, an effective amount of a compound in a formulation for treating a disease, disorder, and / or condition is an amount that relieves, ameliorate, alleviates, inhibits, prevents, delays the onset of, reduces the severity of, and / or reduces the incidence of one or more symptoms or characteristics of the disease, disorder, and / or condition. In some embodiments, a therapeutically effective amount is administered in a single dose: in some embodiments, multiple unit doses are required to deliver a therapeutically effective amount.

[0097] Treat: As used herein, the terms "treat", "treatment" or "treating" refer to any method used to partially or completely alleviate, ameliorate, alleviate, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition. Treatment may be administered to subjects who do not show signs of the disease, disorder, and / or condition. In some embodiments, treatment may be administered to subjects who show only early signs of the disease, disorder, and / or condition, e.g., to reduce the risk of developing pathologies associated with the disease, disorder, and / or condition.

[0098] Unsaturated: The term "unsaturated," as used herein, means that a moiety has one or more units of unsaturation.

[0099] Wild-type: As used herein, the term "wild-type" has its art-recognized meaning and refers to an entity that has structure and / or activity as found in nature in a "normal" (as opposed to mutant, diseased, altered, etc.) state or situation. One of skill in the art will understand that wild-type genes and polypeptides often exist in multiple alternative forms (e.g., alleles).

[0100] As one of skill in the art will understand, the methods and compositions described herein relating to provided compounds (e.g., oligonucleotides) also generally apply to pharma- ceutically acceptable salts of such compounds.

[0101] Description of Specific Embodiments Oligonucleotides are useful in a variety of therapeutic, diagnostic, and research applications. The use of naturally occurring nucleic acids is limited, for example, by their susceptibility to endo- and exo-nucleases. Therefore, various synthetic counterparts have been developed to circumvent these drawbacks and / or further improve various properties and activities. These include, inter alia, synthetic oligonucleotides that contain chemical modifications, such as base modifications, sugar modifications, backbone modifications, etc., that make these molecules less susceptible to degradation and improve other properties and / or activities.

[0102] From a structural standpoint, modifications to the internucleotide linkages can introduce chirality, and certain properties and activities can be influenced by the configuration of the bound phosphorus atoms of the oligonucleotide, such as binding affinity, sequence-specific binding to complementary RNA, stability against nucleases, activity, delivery, pharmacokinetics, etc., among others, can be influenced by the chirality of the backbone bound phosphorus atoms.

[0103] In particular, the present disclosure utilizes techniques to control various structural elements (e.g., sugar modifications and patterns thereof, nucleobase modifications and patterns thereof, modified internucleotide linkages and patterns thereof, stereochemistry of linked phosphorus and patterns thereof, additional chemical moieties (moieties not normally present in an oligonucleotide chain) and patterns thereof, etc.). With the ability to fully control the structural elements of the oligonucleotides, the present disclosure provides oligonucleotides with improved and / or novel properties and / or activities for various applications, e.g., as therapeutic agents, probes, etc. For example, in some embodiments, the provided oligonucleotides and compositions thereof are particularly potent for editing target adenosines in target nucleic acids to correct G to A mutations, in some embodiments, by converting A to I.

[0104] In some embodiments, the oligonucleotide is a 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, The nucleic acid comprises a sequence identical to or completely or substantially complementary to 57, 58, 59, 60, typically 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60 or more consecutive bases. In some embodiments, the nucleic acid is a target nucleic acid comprising one or more target adenosines. In some embodiments, the target nucleic acid comprises no more than one target adenosine. In some embodiments, the oligonucleotide may hybridize to the target nucleic acid. In some embodiments, such hybridization promotes modification of A (eg, conversion of A to I) in a nucleic acid or product thereof, for example, by ADAR1, ADAR2, etc.

[0105] In some embodiments, the present disclosure provides an oligonucleotide comprising: A first domain; and Second Domain Including, a base sequence of the first domain is complementary to a first portion of a base sequence of a target nucleic acid; the base sequence of the second domain is complementary to a second portion of the base sequence of the target nucleic acid; An oligonucleotide is provided in which a first portion and a second portion of the base sequence of a target nucleic acid are separated by a gap.

[0106] In some embodiments, the first domain is capable of hybridizing to a first portion of the target nucleic acid. In some embodiments, the second domain is capable of hybridizing to a second portion of the target nucleic acid. In some embodiments, the first portion and the second portion are separated by a gap. In some embodiments, the present disclosure provides an oligonucleotide comprising: A first domain; and Second Domain Including, the first domain is characterized by being capable of forming a duplex with a first portion of a target nucleic acid; the second domain is characterized as being capable of forming a duplex with a second portion of the target nucleic acid; An oligonucleotide is provided in which a first portion and a second portion of the base sequence of a target nucleic acid are separated by a gap.

[0107] In some embodiments, the length of the first domain is about or at least about 5-100, 5-50, 5-40, 5-30, 5-20, 10-100, 10-50, 10-40, 10-30, 10-20, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases. In some embodiments, the length of the second domain is about or at least about 5-100, 5-50, 5-40, 5-30, 5-20, 10-100, 10-50, 10-40, 10-30, 10-20, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases. In some embodiments, the length of the gap is about or at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, 60, 70, 80, 90, or 100 nucleobases. In some embodiments, the length of the gap is about or at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, 60, 70, 80, 90, or 100 nucleobases. In some embodiments, the gap length is about 2-100,000, 2-50,000, 2-10,000, 2-5,000, 3-100,000, 3-50,000, 3-10,000, 3-5,000, 4-100,000, 4-50,000, 4-10,000, 4-5,000, 5-100,000, 5-50,000, 5-10,000, 5-5,000, 10-1 00,000, 10 to 50,000, 10 to 10,000, 10 to 5,000, 20 to 100,000, 20 to 50,000, 20 to 10,000, 20 to 5,000, 50 to 100,000, 50 to 50,000, 50 to 10,000, 50 to 5,000, 100 to 100,000, 100 to 50,000, 100 to 10,000, or 100 to 5,000 nucleic acid bases.

[0108] In some embodiments, the oligonucleotide has a base sequence that is or includes about 10-40, about 15-40, about 20-40, or at least about 27, at least about 28, at least about 29, at least about 30, at least about 31, at least about 32, at least about 33, at least about 34 consecutive bases of an oligonucleotide or nucleic acid disclosed in a table, or a sequence complementary to a target RNA sequence gene, transcript, etc. disclosed herein, and each T may be optionally and independently replaced with a U, or vice versa. In some embodiments, the disclosure provides an oligonucleotide or oligonucleotide composition as disclosed herein (e.g., in a table). In some embodiments, the base sequence of the oligonucleotide is or includes a sequence that shares about 50%-100%, e.g., about or at least about 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% identity with the first domain of an oligonucleotide in a table. Alternatively or additionally, in some embodiments, the base sequence of the oligonucleotide is or comprises a sequence that shares about 50%-100%, e.g., about or at least about 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% identity with the second domain of the oligonucleotide in the table. In some embodiments, the base sequence of the oligonucleotide is or comprises a sequence that shares about 50%-100%, e.g., about or at least about 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% identity with the oligonucleotide in the table.

[0109] In some embodiments, the oligonucleotide is a single-stranded oligonucleotide for site-specific editing of a nucleoside (e.g., a targeted adenosine) in a target nucleic acid (e.g., RNA).

[0110] As described herein, an oligonucleotide may contain one or more modified internucleotide linkages (non-natural phosphate linkages). In some embodiments, the modified internucleotide linkage is a chiral internucleotide linkage where the linking phosphorus is chiral. In some embodiments, the modified internucleotide linkage is a phosphorothioate internucleotide linkage. In some embodiments, an oligonucleotide comprises one or more negatively charged internucleotide linkages (e.g., phosphorothioate internucleotide linkages, natural phosphate linkages, etc.). In some embodiments, an oligonucleotide comprises one or more non-negatively charged internucleotide linkages. In some embodiments, an oligonucleotide comprises one or more neutral internucleotide linkages.

[0111] In some embodiments, the oligonucleotide is chirally controlled. In some embodiments, the oligonucleotide is chirally pure (or "stereically pure", "stereochemically pure"), and the oligonucleotide exists as a single stereoisomeric form (often a single diastereoisomeric (or "diastereomeric") form, since multiple chiral centers may exist in the oligonucleotide, for example, at the bond phosphorus, sugar, carbon, etc.). As will be understood by those skilled in the art, chirally pure oligonucleotides are separated from other stereoisomeric forms (to the extent that some impurities may exist, since chemical and biological processes, selectivity and / or purification, etc., are rarely, if ever, absolutely perfect). In chirally pure oligonucleotides, each chiral center is independently defined with respect to its configuration (for chirally pure oligonucleotides, each internucleotide bond is independently stereorestricted or chiral controlled). In contrast to chirally controlled and chirally pure oligonucleotides containing sterically restricted linking phosphorus, racemic (or "stereorandom", "chirally uncontrolled") oligonucleotides containing chiral linking phosphorus, derived for example from conventional phosphoramidite oligonucleotide synthesis without stereochemical control during the coupling step in combination with conventional sulfurization (producing stereorandom phosphorothioate internucleotide linkages), refer to an irregular mixture of various stereoisomers (typically diastereoisomers (or "diastereomers"), since there are multiple chiral centers in the oligonucleotide; derived for example from conventional oligonucleotide preparations using reagents that contain no chiral elements other than those at the nucleotides and linking phosphorus). For example, in terms of A*A*A (where * is a phosphorothioate internucleotide linkage (containing a chiral linking phosphorus)), the preparation of racemic oligonucleotides can result in the production of four diastereomers [2 2= 4, considering two chiral linking phosphorus, each of which can exist in either of two configurations (Sp or Rp): A*SA*SA, A*SA*RA, A*RA*SA, and A*RA*RA (*S represents the phosphorothioate internucleotide linkage of Sp, and *R represents the phosphorothioate internucleotide linkage of Rp). With respect to a chirally pure oligonucleotide (e.g., A*SA*SA), the oligonucleotide exists in a single stereoisomeric form, and the oligonucleotide is separated from other stereoisomers (e.g., diastereomers A*SA*RA, A*RA*SA, and A*RA*RA).

[0112] In some embodiments, the oligonucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more sterically irregular internucleotide linkages (a mixture of Rp and Sp linked phosphorus at the internucleotide linkage, e.g., resulting from conventional, non-chiral controlled oligonucleotide synthesis). In some embodiments, the oligonucleotide comprises one or more (e.g., 1-60, 1-50, 1-40, 1-30, 1-25, 1-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 , 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, or more) chirally controlled internucleotide linkages (Rp or Sp linked phosphorus at the internucleotide linkage, e.g., derived from chirally controlled oligonucleotide synthesis). In some embodiments, the internucleotide linkages are phosphorothioate internucleotide linkages. In some embodiments, the internucleotide linkages are stereorandom phosphorothioate internucleotide linkages. In some embodiments, the internucleotide linkages are chirally controlled phosphorothioate internucleotide linkages.

[0113] In particular, the present disclosure provides techniques for preparing chiral controlled (and in some embodiments, stereochemically pure) oligonucleotides. In some embodiments, the oligonucleotides are stereochemically pure. In some embodiments, the oligonucleotides of the present disclosure are about 5%-100%, 10%-100%, 20%-100%, 30%-100%, 40%-100%, 50%-100%, 60%-100%, 70%-100%, 80%-100%, 90%-100%, 95%-100%, 50%-90%, or about 5%, 10%, 15%, 20%, 25%, 30%, 35%. , 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% stereochemically pure.

[0114] In some embodiments, the present disclosure provides various oligonucleotide compositions.In some embodiments, the oligonucleotide compositions are sterically irregular or chirality uncontrolled.In some embodiments, there are no chirality controlled internucleotide bonds in the oligonucleotides of the provided compositions.In some embodiments, the internucleotide bonds of the oligonucleotides in the compositions comprise one or more chirality controlled internucleotide bonds (e.g., chirality controlled oligonucleotide compositions).

[0115] In some embodiments, the oligonucleotide composition comprises a plurality of oligonucleotides sharing a common base sequence, where one or more internucleotide linkages in the oligonucleotides are chiral controlled and one or more internucleotide linkages are sterically irregular (not chiral controlled). In some embodiments, the oligonucleotide composition comprises a plurality of oligonucleotides sharing a common base sequence, where each internucleotide linkage in the oligonucleotides that includes a chiral linking phosphorus is independently a chiral controlled internucleotide linkage. In some embodiments, the oligonucleotides share the same base sequence and the same base and sugar modifications. In some embodiments, the oligonucleotide composition comprises oligonucleotides of the same composition, where one or more internucleotide linkages are chiral controlled and one or more internucleotide linkages are sterically irregular (not chiral controlled). In some embodiments, the oligonucleotide composition comprises oligonucleotides of the same composition, where each internucleotide linkage in the oligonucleotides that includes a chiral linking phosphorus is independently a chiral controlled internucleotide linkage. In some embodiments, at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% or all of the oligonucleotides of the common base sequence are multiple oligonucleotides.

[0116] In some embodiments, the disclosure provides techniques for preparing, evaluating, and / or utilizing the provided oligonucleotides and compositions thereof.

[0117] As used in this disclosure, in some embodiments, "one or more" is 1 to 200, 1 to 150, 1 to 100, 1 to 90, 1 to 80, 1 to 70, 1 to 60, 1 to 50, 1 to 40, 1 to 30, or is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60. In some embodiments, "one or more" is 1. In some embodiments, "one or more" is 2. In some embodiments, "one or more" is 3. In some embodiments, "one or more" is 4. In some embodiments, "one or more" is 5. In some embodiments, "one or more" is 6. In some embodiments, "one or more" is 7. In some embodiments, "one or more" is 8. In some embodiments, "one or more" is 9. In some embodiments, "one or more" is 10. In some embodiments, "one or more" is at least 1. In some embodiments, "one or more" is at least 2. In some embodiments, "one or more" is at least 3. In some embodiments, "one or more" is at least 4. In some embodiments, "one or more" is at least 5. In some embodiments, "one or more" is at least 6. In some embodiments, "one or more" is at least 7. In some embodiments, "one or more" is at least 8. In some embodiments, "one or more" is at least 9. In some embodiments, "one or more" is at least ten.

[0118] As used in this disclosure, in some embodiments, "at least one" means one or more.

[0119] For example, variables (e.g., R, R LVarious embodiments are described with respect to variables (e.g., R), L, etc. An embodiment described with respect to a variable (e.g., R) generally includes all variables that may be such a variable (e.g., R′, R″, R L , R L1 etc.)

[0120] Oligonucleotides In particular, the present disclosure provides oligonucleotides of various designs, which may include various nucleobases and their patterns, sugars and their patterns, internucleotide linkages and their patterns, and / or additional chemical moieties and their patterns as described in the present disclosure. In some embodiments, the provided oligonucleotides can induce A to I editing in a target nucleic acid. In some embodiments, the oligonucleotides of the present disclosure are single-stranded oligonucleotides capable of site-specific editing (A to I conversion) of adenosines in a target RNA sequence.

[0121] In some embodiments, the oligonucleotide is of suitable length and sequence complementary to specifically hybridize with the target nucleic acid. In some embodiments, the oligonucleotide is of sufficient length and sufficiently complementary to the target nucleic acid to distinguish the target nucleic acid from other nucleic acids to reduce off-target effects. In some embodiments, the oligonucleotide is short enough to facilitate delivery and reduce manufacturing complexity and / or cost, while maintaining the desired properties and activity (e.g., adenosine editing).

[0122] In some embodiments, the oligonucleotides are about 10 to 200 (e.g., about 10 to 20, 10 to 30, 10 to 40, 10 to 50, 10 to 60, 10 to 70, 10 to 80, 10 to 90, 10 to 100, 10 to 120, 10 to 150, 20 to 30, 20 to 40, 20 to 50, 20 to 60, 20 to 70, 20 to 80, 20 to 90, 20 to 100, 20 to 120, 20 to 150, 20 to 200, 25 to 30, 25 to 40, 25 to 50, 25-60, 25-70, 25-80, 25-90, 25-100, 25-120, 25-150, 25-200, 30-40, 30-50, 30-60, 30-70, 30-80, 30-90, 30-100, 30-120, 30-150, 30-200, 10, 20, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 45, 50, 60, etc.) nucleic acid bases in length. In some embodiments, the oligonucleotide base sequence is about 10-60 nucleic acid bases in length. In some embodiments, the base sequence is about 15-50 nucleic acid bases in length. In some embodiments, the base sequence is about 15 to about 35 nucleobases in length. In some embodiments, the base sequence is about 25 to about 34 nucleobases in length. In some embodiments, the base sequence is about 26 to about 35 nucleobases in length. In some embodiments, the base sequence is about 27 to about 32 nucleobases in length. In some embodiments, the base sequence is about 29 to about 35 nucleobases in length. In some embodiments, the base sequence is about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 nucleobases in length. In some embodiments, the base sequence is 35 nucleobases in length or at least 35 nucleobases in length. In some embodiments, the base sequence is 34 nucleobases in length or at least 34 nucleobases in length. In some embodiments, the base sequence is 33 nucleobases in length or at least 33 nucleobases in length.In some embodiments, the base sequence is 32 nucleobases long or at least 32 nucleobases long. In some embodiments, the base sequence is 31 nucleobases long or at least 31 nucleobases long. In some embodiments, the base sequence is 30 nucleobases long or at least 30 nucleobases long. In some embodiments, the base sequence is 29 nucleobases long or at least 29 nucleobases long. In some embodiments, the base sequence is 28 nucleobases long or at least 28 nucleobases long. In some embodiments, the base sequence is 27 nucleobases long or at least 27 nucleobases long. In some embodiments, the base sequence is 26 nucleobases long or at least 26 nucleobases long. In some embodiments, the base sequence of the complementary portions in the duplex is about or at least about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 16, 27, 28, 29, 30, 31, 32, 33, 34, 35 or more nucleobases in length. In some embodiments, it is at least 10 nucleobases in length. In some embodiments, it is at least 11 nucleobases in length. In some embodiments, it is at least 12 nucleobases in length. In some embodiments, it is at least 13 nucleobases in length. In some embodiments, it is at least 14 nucleobases in length. In some embodiments, it is at least 16 nucleobases in length. In some embodiments, it is at least 16 nucleobases in length. In some embodiments, it is at least 17 nucleobases in length. In some embodiments, it is at least 18 nucleobases in length. In some embodiments, it is at least 19 nucleobases in length. In some embodiments, it is at least 20 nucleobases long. In some embodiments, it is at least 21 nucleobases long. In some embodiments, it is at least 22 nucleobases long. In some embodiments, it is at least 23 nucleobases long. In some embodiments, it is at least 24 nucleobases long. In some embodiments, it is at least 25 nucleobases long.In particular, the present disclosure provides oligonucleotides of similar or better properties and / or similar or higher activity but of shorter length compared to previously reported adenosine-edited oligonucleotides.

[0123] In some embodiments, the base sequence of the oligonucleotide or a fragment thereof (e.g., the first domain, the second domain, etc.) is complementary to the base sequence of the target nucleic acid or a fragment thereof (e.g., the first portion, the second portion, etc.) with 0 to 10 (e.g., 0 to 1, 0 to 2, 0 to 3, 0 to 4, 0 to 5, 0 to 6, 0 to 7, 0 to 8, 0 to 9, 0 to 10, 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 2 to 3, 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 2 to 10, 3 to 4, 3 to 5, 3 to 6, 3 to 7, 3 to 8, 3 to 9, 3 to 10, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.) mismatches that are not Watson-Crick base pairs (AT, AU, and CG). In some embodiments, there are zero mismatches. In some embodiments, there is one mismatch. In some embodiments, there are two mismatches. In some embodiments, there are three mismatches. In some embodiments, there are four mismatches. In some embodiments, there are five mismatches. In some embodiments, there are six mismatches. In some embodiments, there are seven mismatches. In some embodiments, there are eight mismatches. In some embodiments, there are nine mismatches. In some embodiments, there are ten mismatches. In some embodiments, the complementarity (e.g., complementarity between an oligonucleotide or a fragment thereof and a target nucleic acid or a fragment thereof (e.g., between a first domain and a first portion, between a second domain and a second portion, etc.)) is about 50% to 100% (e.g., about 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65%~100%, 70%~80%, 70%~85%, 70%~90%, 70%~95%, 70%~100%, 75%~80%, 75%~85%, 75%~90%, 75%~95%, 75%~100%, 80%~85%, 80%~90%, 80%~95%, 80%~100%, 85%~90%, 85%~95%, 85%~100%, 90%~95%, 90%~100%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, etc.In some embodiments, the complementarity is at least about 60%. In some embodiments, the complementarity is at least about 65%. In some embodiments, the complementarity is at least about 70%. In some embodiments, the complementarity is at least about 75%. In some embodiments, the complementarity is at least about 80%. In some embodiments, the complementarity is at least about 85%. In some embodiments, the complementarity is at least about 90%. In some embodiments, the complementarity is at least about 95%. In some embodiments, the complementarity is 100%. In some embodiments, the complementarity is 100% over the entire length of the oligonucleotide, except for the nucleoside opposite the target nucleoside (e.g., adenosine). Typically, the complementarity is based on Watson-Crick base pairs AT, AU, and CG. One of skill in the art will understand that when assessing the complementarity of two sequences of different lengths (e.g., a provided oligonucleotide and a target nucleic acid), the complementarity may be appropriately based on the length of the shorter sequence and / or the maximum complementarity between the two sequences. In many embodiments, the oligonucleotide and the target nucleic acid are sufficiently complementary such that modifications are selectively directed to the target adenosine site.

[0124] In some embodiments, one or more mismatches are independently perturbed. In some embodiments, each mismatch is perturbed. In some embodiments, there are 0 to 10 perturbations (e.g., 0 to 1, 0 to 2, 0 to 3, 0 to 4, 0 to 5, 0 to 6, 0 to 7, 0 to 8, 0 to 9, 0 to 10, 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 2 to 3, 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 2 to 10, 3 to 4, 3 to 5, 3 to 6, 3 to 7, 3 to 8, 3 to 9, 3 to 10, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.). In some embodiments, this number is 0. In some embodiments, this number is 1. In some embodiments, this number is 2. In some embodiments, this number is 3. In some embodiments, this number is 4. In some embodiments, this number is 5. In some embodiments, the wobble is GU, IA, GA, IU, IC, IT, AA, or inverted AT. In some embodiments, the wobble is GU, IA, GA, IU, or IC. In some embodiments, an IC can be considered a match when an I is the nucleoside immediately 3' to the nucleoside opposite the target nucleoside. In some embodiments, a base that forms a wobble pair (e.g., U, which can form a GU wobble) can replace a base that forms a match pair (e.g., C, which matches G) to provide an oligonucleotide with editing activity.

[0125] In some embodiments, the oligonucleotide and target nucleic acid duplexes contain one or more bulges, each independently containing one or more mismatches that are not wobble. In some embodiments, there are 0 to 10 bulges (e.g., 0 to 1, 0 to 2, 0 to 3, 0 to 4, 0 to 5, 0 to 6, 0 to 7, 0 to 8, 0 to 9, 0 to 10, 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 2 to 3, 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 2 to 10, 3 to 4, 3 to 5, 3 to 6, 3 to 7, 3 to 8, 3 to 9, 3 to 10, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.). In some embodiments, this number is 0. In some embodiments, this number is 1. In some embodiments, this number is 2. In some embodiments, this number is 3. In some embodiments, this number is 4. In some embodiments, this number is 5.

[0126] In some embodiments, the distance between the two mismatches, the mismatch and one or both ends of the oligonucleotide (or portions thereof, e.g., the first domain, the second domain, the first subdomain, the second subdomain, the third subdomain), and / or the nucleoside opposite the mismatch and the target adenosine, is independently 0 to 50, 0 to 40, 0 to 30, 0 to 25, 0 to 20, 0 to 15, 0 to 10 (e.g., 0 to 1, 0 to 2, 0 to 3, 0 to 4, 0 to 5, 0 to 6, 0 to 7, 0 to 8, 0 to 9, 0 to 10, 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to It can be 6, 1-7, 1-8, 1-9, 1-10, 2-3, 2-4, 2-5, 2-6, 2-7, 2-8, 2-9, 2-10, 3-4, 3-5, 3-6, 3-7, 3-8, 3-9, 3-10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 nucleobases (not including mismatches, terminal nucleosides, and nucleosides opposite the target adenosine). In some embodiments, this number is , 0 to 30. In some embodiments, this number is 0 to 20. In some embodiments, this number is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In some embodiments, the distance between two mismatches is 0 to 20. In some embodiments, the distance between two mismatches is 1 to 10. In some embodiments, the distance between a mismatch and the 5'-terminal nucleoside of an oligonucleotide is 0 to 20. In some embodiments, the distance between a mismatch and the 5'-terminal nucleoside of an oligonucleotide is 0 to 20. The distance between the 5'-terminal nucleoside of the oligonucleotide is 5 to 20. In some embodiments, the distance between the mismatch and the 3'-terminal nucleoside of the oligonucleotide is 0 to 40. In some embodiments, the distance between the mismatch and the 3'-terminal nucleoside of the oligonucleotide is 5 to 20. In some embodiments, the distance between the mismatch and the nucleoside opposite the target adenosine is 0 to 20. In some embodiments, the distance between the mismatch and the nucleoside opposite the target adenosine is 1 to 10.In some embodiments, the number of nucleobases for the distance is 0. In some embodiments, it is 1. In some embodiments, it is 2. In some embodiments, it is 3. In some embodiments, it is 4. In some embodiments, it is 5. In some embodiments, it is 6. In some embodiments, it is 7. In some embodiments, it is 8. In some embodiments, it is 9. In some embodiments, it is 10. In some embodiments, it is 11. In some embodiments, it is 12. In some embodiments, it is 13. In some embodiments, it is 14. In some embodiments, it is 15. In some embodiments, it is 16. In some embodiments, it is 17. In some embodiments, it is 18. In some embodiments, it is 19. In some embodiments, it is 20. In some embodiments, the mismatch is at the end (e.g., at the 5'-end or 3'-end) of the first domain, the second domain, the first subdomain, the second subdomain, or the third subdomain. In some embodiments, the mismatch is at the nucleoside opposite the targeted adenosine.

[0127] In some embodiments, provided oligonucleotides are capable of inducing an adenosine edit (e.g., A to I conversion) in a target nucleic acid and have a base sequence that consists of, comprises, or comprises a portion (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or more consecutive bases) of the base sequence of an oligonucleotide disclosed herein, wherein each T can be independently replaced with a U, and vice versa, and the oligonucleotide comprises at least one non-naturally occurring modification of the base, sugar, and / or internucleotide linkage.

[0128] In some embodiments, the oligonucleotide provided comprises one or more carbohydrate moieties. In some embodiments, the oligonucleotide provided comprises one or more GalNAc moieties. In some embodiments, the oligonucleotide provided comprises one or more targeting moieties. Non-limiting examples of such additional chemical moieties that can be conjugated to oligonucleotide chains are described herein.

[0129] In some embodiments, the provided oligonucleotides can induce the correction of G to A mutations in a target sequence or its product. In some embodiments, the correction of G to A mutations is or includes the conversion of A to I, which can be read as G during translation or other biological processes. In some embodiments, the provided oligonucleotides can induce the correction of G to A mutations in a target sequence or its product through ADAR-mediated deamination. In some embodiments, the provided oligonucleotides can induce the correction of G to A mutations in a target sequence or its product through ADAR-mediated deamination by recruiting endogenous ADARs (e.g., in target cells) and promoting ADAR-mediated deamination. However, the present disclosure is not limited to any particular mechanism nevertheless. In some embodiments, the present disclosure provides oligonucleotides, compositions, methods, etc. that can operate through double-stranded RNA interference, single-stranded RNA interference, RNase H-mediated knockdown, steric hindrance of translation, ADAR-mediated deamination, or a combination of two or more such mechanisms.

[0130] In some embodiments, the oligonucleotide comprises a structural element or portion thereof described herein (e.g., in a table). In some embodiments, the oligonucleotide has a base sequence that comprises a base sequence (or portion thereof), where each T can be independently replaced with a U, a pattern of chemical modifications (or portions thereof) disclosed herein (e.g., in a table or figure) or otherwise disclosed herein, and / or the format of the oligonucleotide. In some embodiments, such an oligonucleotide can induce correction of a G to A mutation in a target sequence or its product.

[0131] In particular, the provided oligonucleotides can hybridize to their target nucleic acid (e.g., pre-mRNA, mature mRNA, etc.). In some embodiments, the oligonucleotides can hybridize to the nucleic acid of the target RNA sequence at any stage of RNA processing, including, but not limited to, pre-mRNA or mature mRNA. In some embodiments, the oligonucleotides can hybridize to any element of the oligonucleotide nucleic acid or its complement, including, but not limited to, a promoter region, an enhancer region, a transcription termination region, a translation initiation signal, a translation termination signal, a coding region, a non-coding region, an exon, an intron, an intron / exon or an exon / intron junction, a 5'UTR, or a 3'UTR.

[0132] In some embodiments, the oligonucleotide hybridizes to two or more variants of a transcript derived from the sense strand of a target site (eg, a target sequence).

[0133] In some embodiments, the provided oligonucleotides contain an increased level of one or more isotopes. In some embodiments, the provided oligonucleotides are labeled, for example, with one or more isotopes of one or more elements (e.g., hydrogen, carbon, nitrogen, etc.). In some embodiments, the provided oligonucleotides in the provided compositions (e.g., multiple oligonucleotides of the composition) include base modifications, sugar modifications, and / or internucleotide linkage modifications, and the oligonucleotides contain enriched levels of deuterium. In some embodiments, the provided oligonucleotides are enriched with deuterium (-) at one or more positions. 1 H- 2 In some embodiments, one or more of the oligonucleotide strands or any moieties conjugated to the oligonucleotide strands (e.g., targeting moieties, etc.) are labeled with 1 H, 2 substituted with H. Such oligonucleotides may be used in the compositions and methods described herein.

[0134] In some embodiments, the oligonucleotide comprises one or more modified nucleobases, one or more modified sugars, and / or one or more modified internucleotide linkages, as described herein. In some embodiments, the oligonucleotide comprises, for example, about 5%-100%, about 10%-100%, 20-100%, 30%-100%, 40%-100%, 50%-80%, 50%-85%, 50%-90%, 50%-95%, 60%-80%, 60%-85%, 60%-90%, 60%-95%, 60%-100%, 65%-80%, 65%-85%, 65%-90%, 65%-95%, 65%-100%, 70%-80%, 70%-85%, 70%-90 ... and / or 100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.

[0135] In some embodiments, the oligonucleotide comprises one or more modified sugars. In some embodiments, the oligonucleotide comprises from about 1 to 50 (e.g., from about 5, 6, 7, 8, 9, or 10 to about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, etc., such as about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, etc.) modified sugars. In some embodiments, the oligonucleotide comprises about 1 to 50 (e.g., about 5, 6, 7, 8, 9, or 10 to about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, etc., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, etc.) modified sugars having 2'-F modifications.In some embodiments, the oligonucleotide has about 2-50 (e.g., about 2, 3, 4, 5, 6, 7, 8, 9, or 10 to about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, etc., 2-40, 2-30, 2-25, 2-20, 2-15, 2-10, 3-40, 3-30, 3-25, 3-20, 3-15, 3-10, 4-40, 4- 30, 4~25, 4~20, 4~15, 4~10, 5~40, 5~30, 5~25, 5~20, 5~15, 5~10, 6~40, 6~30, 6~25, 6~20, 6~15, 6~10, 7~40, 7~30, 7~25, 7~20, 7~15, 7~10, 8~40, 8~30, 8~25, 8~20, 8~15 , 8-10, 9-40, 9-30, 9-25, 9-20, 9-15, 9-10, 10-40, 10-30, 10-25, 10-20, 10-15, about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 consecutive modified sugars. In some embodiments, the oligonucleotide comprises two consecutive 2'-F modified sugars. In some embodiments, the oligonucleotide comprises three consecutive 2'-F modified sugars. In some embodiments, the oligonucleotide comprises four consecutive 2'-F modified sugars. In some embodiments, the oligonucleotide comprises five consecutive 2'-F modified sugars. In some embodiments, the oligonucleotide comprises six consecutive 2'-F modified sugars. In some embodiments, the oligonucleotide comprises seven consecutive 2'-F modified sugars. In some embodiments, the oligonucleotide comprises 8 contiguous 2'-F modified sugars. In some embodiments, the oligonucleotide comprises 9 contiguous 2'-F modified sugars. In some embodiments, the oligonucleotide comprises 10 contiguous 2'-F modified sugars.In some embodiments, an oligonucleotide comprises two or more 2'-F modified sugar blocks, where each sugar in a 2'-F modified sugar block is independently a 2'-F modified sugar. In some embodiments, each 2'-F modified sugar block independently comprises or consists of 2, 3, 4, 5, 6, 7, 8, 9, or 10 contiguous 2'-F modified sugars as described herein. In some embodiments, two contiguous 2'-F modified sugar blocks are independently separated by a separation block, where the separation block independently comprises one or more sugars that are not 2'-F modified sugars. In some embodiments, the oligonucleotide comprises one or more (e.g., 1-20, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 2-20, 3-15, 4-15, 5-15, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.) 2'-F blocks and one or more (e.g., 1-20, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 2-20, 3-15, 4-15, 5-15, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.) separation blocks. In some embodiments, the first domain is one or more (e.g., 1-20, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 2-20, 3-15, 4-15, 5-15, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.) and one or more (e.g., 1-20, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 2-20, 3-15, 4-15, 5-15, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.) separation blocks. In some embodiments, each first domain block attached to a first domain 2'-F block is a separation block. In some embodiments, each first domain block attached to a first domain separation block is a first domain 2'-F block. In some embodiments, each sugar in a separation block is independently not 2'-F modified.In some embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more) or all sugars in a separation block are independently not 2'-F modified. In some embodiments, a separation block comprises one or more bicyclic sugars (e.g., LNA sugars, cEt sugars, etc.), and / or one or more 2'-OR modified sugars (wherein R is an optionally substituted C. 1~6 In some embodiments, the separation block comprises one or more 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the two or more 2'-F unmodified sugars are contiguous. In some embodiments, the two or more 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the separation block is comprised of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more) 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the separation block comprises two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more) contiguous 2'-ORF modified sugars (wherein R is an optionally substituted C 1~6In some embodiments, each 2'-OR modified sugar is independently a 2'-OMe sugar or a 2'-MOE sugar. In some embodiments, each 2'-OR modified sugar is independently a 2'-OMe sugar. In some embodiments, each 2'-OR modified sugar is independently a 2'-MOE sugar. In some embodiments, the separation block comprises one or more 2'-F modified sugars. In some embodiments, the 2'-F modified sugars in the separation block are not adjacent to one another. In some embodiments, the separation block does not comprise a 2'-F modified sugar. In some embodiments, each sugar in the separation block is independently a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, each sugar in each separation block is independently a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, each sugar in the separation block is independently a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, each sugar in each separation block is independently a 2'-OR modified sugar, where R is an optionally substituted C 1~6 aliphatic). In some embodiments, each sugar in a separation block is independently a 2'-OMe or 2'-MOE modified sugar. In some embodiments, each sugar in each separation block is independently a 2'-OMe or 2'-MOE modified sugar. In some embodiments, each sugar in a separation block is independently a 2'-OMe modified sugar. In some embodiments, each sugar in a separation block is independently a 2'-MOE modified sugar. In some embodiments, a separation block comprises a 2'-OMe sugar and a 2'-MOE modified sugar. In some embodiments, each 2'-F block and each separation block independently comprises 1, 2, 3, 4, or 5 nucleosides. In some embodiments, each 2'-F block and each separation block independently comprises 1, 2, or 3 nucleosides.

[0136] In some embodiments, between about 10% and 100%, 20% and 100%, 30% and 100%, 40% and 100%, 50% and 80%, 50% and 85%, 50% and 90%, 50% and 95%, 60% and 80%, 60% and 85%, 60% and 90%, 60% and 95%, 60% and 100%, 65% and 80%, 65% and 85%, 65% and 90%, 65% and 95%, 65% and 100%, 70% and 80%, 70% and 85%, 70% and 90%, 70% and 95% of all sugars. %, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% is a modified sugar. In some embodiments, between about 10% and 100%, 20% and 100%, 30% and 100%, 40% and 100%, 50% and 80%, 50% and 85%, 50% and 90%, 50% and 95%, 60% and 80%, 60% and 85%, 60% and 90%, 60% and 95%, 60% and 100%, 65% and 80%, 65% and 85%, 65% and 90%, 65% and 95%, 65% and 100%, 70% and 80%, 70% and 85%, 70% and 90%, 70% and 95%, 70% and 100%, 75% and 8 ... 0%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% may be 2'-F modified sugars, 2'-OR modified sugars (wherein R is an optionally substituted C 1~6aliphatic), and bicyclic sugars (e.g., LNA sugars, cEt sugars, etc.). In some embodiments, the percentage is about or at least about 30%. In some embodiments, the percentage is about or at least about 40%. In some embodiments, the percentage is about or at least about 50%. In some embodiments, the percentage is about or at least about 60%. In some embodiments, the percentage is about or at least about 70%. In some embodiments, the percentage is about or at least about 80%. In some embodiments, the percentage is about or at least about 90%. In some embodiments, the percentage is about or at least about 95%.

[0137] In some embodiments, between about 10% and 100%, 20% and 100%, 30% and 100%, 40% and 100%, 50% and 80%, 50% and 85%, 50% and 90%, 50% and 95%, 60% and 80%, 60% and 85%, 60% and 90%, 60% and 95%, 60% and 100%, 65% and 80%, 65% and 85%, 65% and 90%, 65% and 95%, 65% and 100%, 70% and 80%, 70% and 85%, 70% and 90%, 70% and 95%, 70% and 100%, 75% and 80%, %, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% refers to 2'-F modified sugars and 2'-OR modified sugars (wherein R is an optionally substituted C 1~6In some embodiments, the modified sugars are independently selected from the group consisting of about 10%-100%, 20-100%, 30%-100%, 40%-100%, 50%-80%, 50%-85%, 50%-90%, 50%-95%, 60%-80%, 60%-85%, 60%-90%, 60%-95%, 60%-100%, 65%-80%, 65%-85%, 65%-90%, 65%-95%, 65%-100%, 70%-80%, 70%-85%, 70%-90%, 70%-95%, 70%-100%, 75%-80%, 75%-85%, %, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% are modified sugars independently selected from 2'-F modified sugars, 2'-OMe modified sugars, and 2'-MOE modified sugars. In some embodiments, the percentage is about or at least about 30%. In some embodiments, the percentage is about or at least about 40%. In some embodiments, the percentage is about or at least about 50%. In some embodiments, the percentage is about or at least about 60%. In some embodiments, the percentage is about or at least about 70%. In some embodiments, the percentage is about or at least about 80%. In some embodiments, the percentage is about or at least about 90%. In some embodiments, the percentage is about or at least about 95%.

[0138] In some embodiments, between about 10%-100%, 20-100%, 30%-100%, 40%-100%, 50%-80%, 50%-85%, 50%-90%, 50%-95%, 60%-80%, 60%-85%, 60%-90%, 60%-95%, 60%-100%, 65%-80%, 65%-85%, 65%-90%, 65%-95%, 65%-100%, 70%-80%, 70%-85%, 70%-90%, 70%-95%, 70%-100%, 75%-80% , 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% are modified sugars independently selected from 2'-F modified sugars and 2'-OMe modified sugars. In some embodiments, the percentage is about or at least about 30%. In some embodiments, the percentage is about or at least about 40%. In some embodiments, the percentage is about or at least about 50%. In some embodiments, the percentage is about or at least about 60%. In some embodiments, the percentage is about or at least about 70%. In some embodiments, the percentage is about or at least about 80%. In some embodiments, the percentage is about or at least about 90%. In some embodiments, the percentage is about or at least about 95%.

[0139] In some embodiments, between about 10% and 100%, 20% and 100%, 30% and 100%, 40% and 100%, 50% and 80%, 50% and 85%, 50% and 90%, 50% and 95%, 60% and 80%, 60% and 85%, 60% and 90%, 60% and 95%, 60% and 100%, 65% and 80%, 65% and 85%, 65% and 90%, 65% and 95%, 65% and 100%, 70% and 80%, 70% and 85%, 70% and 90%, 70% and 95%, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% are 2'-F modified sugars. In some embodiments, the percentage is about or at least about 30%. In some embodiments, the percentage is about or at least about 40%. In some embodiments, the percentage is about or at least about 50%. In some embodiments, the percentage is about or at least about 60%. In some embodiments, the percentage is about or at least about 70%. In some embodiments, the percentage is about or at least about 80%. In some embodiments, the percentage is about or at least about 90%. In some embodiments, the percentage is about or at least about 95%. In some embodiments, 10 or more (e.g., about or at least about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more, such as 10-50, 10-40, 10-30, 10-25, 15-50, 15-40, 15-30, 15-25, 20-50, 20-40, 20-30, 20-25, etc.) sugars are 2'-F modified sugars.In some embodiments, the oligonucleotide comprises two or more (e.g., 2-30, 2-25, 2-20, 2-15, 3-10, 3-30, 3-25, 3-20, 3-15, 3-10, 4-30, 4-25, 4-20, 4-15, 4-10, 5-30, 5-25, 5-20, 5-15, 5-10, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) consecutive 2'-F modified sugars. In some embodiments, the oligonucleotide comprises one or more 2'-F blocks each independently comprising two or more (e.g., 2-30, 2-25, 2-20, 2-15, 3-10, 3-30, 3-25, 3-20, 3-15, 3-10, 4-30, 4-25, 4-20, 4-15, 4-10, 5-30, 5-25, 5-20, 5-15, 5-10, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) consecutive 2'-F modified sugars. In some embodiments, the oligonucleotide comprises two or more 2'-F blocks as described herein separated by one or more separation blocks as described herein. In some embodiments, the 2'-F block has 2, 3, 4, 5, 6, 7, 8, 9, or 10 2'-F modified sugars. In some embodiments, the 2'-F block has no more than 2, 3, 4, 5, 6, 7, 8, 9, or 10 2'-F modified sugars. In some embodiments, each sugar in each 2'-F block is a 2'-F modified sugar, and each 2'-F block independently has 2, 3, 4, 5, 6, 7, 8, 9, or 10 2'-F modified sugars. In some embodiments, each sugar in each 2'-F block is a 2'-F modified sugar, and each 2'-F block independently has 2, 3, 4, 5, 6, 7, 8, 9, or 10 2'-F modified sugars. In some embodiments, each sugar in each 2'-F block is a 2'-F modified sugar, and each 2'-F block independently has 10 or fewer 2'-F modified sugars. In some embodiments, each sugar in each 2'-F block is a 2'-F modified sugar, and each 2'-F block independently has 9 or fewer 2'-F modified sugars.In some embodiments, each sugar in each 2'-F block is a 2'-F modified sugar, and each 2'-F block independently has 8 or fewer 2'-F modified sugars. In some embodiments, each sugar in each 2'-F block is a 2'-F modified sugar, and each 2'-F block independently has 7 or fewer 2'-F modified sugars. In some embodiments, each sugar in each 2'-F block is a 2'-F modified sugar, and each 2'-F block independently has 6 or fewer 2'-F modified sugars. In some embodiments, each sugar in each 2'-F block is a 2'-F modified sugar, and each 2'-F block independently has 5 or fewer 2'-F modified sugars. In some embodiments, each sugar in each 2'-F block is a 2'-F modified sugar, and each 2'-F block independently has 4 or fewer 2'-F modified sugars. In some embodiments, each block attached to a 2'-F block is independently a block that does not contain a 2'-F modified sugar. In some embodiments, each block attached to a 2'-F block is independently a block that contains a neutral DNA or RNA sugar, a 2'-OR modified sugar (wherein R is an optionally substituted C). 1~6 In some embodiments, each block attached to the 2'-F block is independently a block comprising a neutral DNA or RNA sugar, a 2'-OMe modified sugar, a 2'-MOE modified sugar, or a bicyclic sugar. In some embodiments, each block attached to the 2'-F block is independently a block comprising a neutral DNA or RNA sugar, a 2'-OMe modified sugar, a 2'-MOE modified sugar. In some embodiments, each nucleoside in the first domain attached to the 2'-F block in the first domain is independently a block comprising a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, each nucleoside in the first domain linked to a 2'-F block in the first domain is independently a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6In some embodiments, each nucleoside in the first domain linked to a 2'-F block in the first domain is independently a 2'-OMe or 2'-MOE modified sugar. In some embodiments, each nucleoside in the second domain linked to a 2'-F block in the second domain is independently a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, each nucleoside in the second domain linked to a 2'-F block in the second domain is independently a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, each nucleoside in the second domain linked to a 2'-F block in the second domain is independently a 2'-OMe or a 2'-MOE modified sugar.

[0140] In some embodiments, between about 10% and 100%, 20% and 100%, 30% and 100%, 40% and 100%, 50% and 80%, 50% and 85%, 50% and 90%, 50% and 95%, 60% and 80%, 60% and 85%, 60% and 90%, 60% and 95%, 60% and 100%, 65% and 80%, 65% and 85%, 65% and 90%, 65% and 95%, 65% and 100%, 70% and 80%, 70% and 85%, 70% and 90%, 70% and 95%, 70% and 100% of all sugars. , 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% is a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6In some embodiments, the total sugars are about 10%-100%, 20-100%, 30%-100%, 40%-100%, 50%-80%, 50%-85%, 50%-90%, 50%-95%, 60%-80%, 60%-85%, 60%-90%, 60%-95%, 60%-100%, 65%-80%, 65%-85%, 65%-90%, 65%-95%, 65%-100%, 70%-80%, 70%-85%, 70%-90%, 70%-95%, 70 ... 00%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% are 2'-OMe or 2'-MOE modified sugars. In some embodiments, the percentage is about or at least about 30%. In some embodiments, the percentage is about or at least about 40%. In some embodiments, the percentage is about or at least about 50%. In some embodiments, the percentage is about or at least about 60%. In some embodiments, the percentage is about or at least about 70%. In some embodiments, the percentage is about or at least about 80%. In some embodiments, the percentage is about or at least about 90%. In some embodiments, the percentage is about or at least about 95%.

[0141] In some embodiments, between about 10% and 100%, 20% and 100%, 30% and 100%, 40% and 100%, 50% and 80%, 50% and 85%, 50% and 90%, 50% and 95%, 60% and 80%, 60% and 85%, 60% and 90%, 60% and 95%, 60% and 100%, 65% and 80%, 65% and 85%, 65% and 90%, 65% and 95%, 65% and 100%, 70% and 80%, 70% and 85%, 70% and 90%, 70% and 95 ... 0%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% are 2'-OMe modified sugars. In some embodiments, the percentage is about or at least about 30%. In some embodiments, the percentage is about or at least about 40%. In some embodiments, the percentage is about or at least about 50%. In some embodiments, the percentage is about or at least about 60%. In some embodiments, the percentage is about or at least about 70%. In some embodiments, the percentage is about or at least about 80%. In some embodiments, the percentage is about or at least about 90%. In some embodiments, the percentage is about or at least about 95%.

[0142] In some embodiments, between about 10% and 100%, 20% and 100%, 30% and 100%, 40% and 100%, 50% and 80%, 50% and 85%, 50% and 90%, 50% and 95%, 60% and 80%, 60% and 85%, 60% and 90%, 60% and 95%, 60% and 100%, 65% and 80%, 65% and 85%, 65% and 90%, 65% and 95%, 65% and 100%, 70% and 80%, 70% and 85%, 70% and 90%, 70% and 95%, 70% and 100% of all sugars. , 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% is a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, the total sugars are about 10%-100%, 20-100%, 30%-100%, 40%-100%, 50%-80%, 50%-85%, 50%-90%, 50%-95%, 60%-80%, 60%-85%, 60%-90%, 60%-95%, 60%-100%, 65%-80%, 65%-85%, 65%-90%, 65%-95%, 65%-100%, 70%-80%, 70%-85%, 70%-90%, 70%-95%, 70%-80%, 70%-85%, 70%-90%, 70%-95%, 70%-95%, 70%-95%, 70%-100%, 70%-80%, 70%-85%, 70%-90%, 70%-95%, 70%-95%, 70%-100%, 70%-80%, 70%-85%, 70%-90%, 70%-95%, 70%-95%, 70%-100%, 70%-80%, 70%-85%, 70%-90%, 70%-95%, 70%-10 ... 0% to 100%, 75% to 80%, 75% to 85%, 75% to 90%, 75% to 95%, 75% to 100%, 80% to 85%, 80% to 90%, 80% to 95%, 80% to 100%, 85% to 90%, 85% to 95%, 85% to 100%, 90% to 95%, 90% to 100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% is a 2'-MOE modified sugar.

[0143] In some embodiments, the first (5' end) one or several (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, 2-3, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 3-4, etc.) nucleosides, and / or The sugars of the last (3'-end) one or several (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, 2-3, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 3-4, etc.) nucleosides are independently modified sugars. In some embodiments, the first one or several sugars are independently modified sugars. In some embodiments, the last one or several sugars are independently modified sugars. In some embodiments, both the first one or several sugars and the last one or several sugars are independently modified sugars. In some embodiments, the modified sugars are independently a 2'-F non-modified sugar (e.g., a bicyclic sugar), a 2'-OR modified sugar (where R is as described herein and is not -H (e.g., an optionally substituted C 1~6 In some embodiments, the modified sugars are independently selected from bicyclic sugars and 2'-OR modified sugars, where R is an optionally substituted C 1~6 In some embodiments, the modified sugar is independently selected from a 2'-OR modified sugar, where R is an optionally substituted C 1~6 In some embodiments, the modified sugars are, independently, 2'-OMe modified sugars and 2'-MOE modified sugars. In some embodiments, the first several sugars are one or more 2'-OR modified sugars (wherein R is an optionally substituted C 1~6In some embodiments, the first several sugars include one or more 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the first several sugars comprise one or more 2'-OMe modified sugars. In some embodiments, the first several sugars comprise one or more 2'-MOE modified sugars. In some embodiments, the first several sugars comprise one or more 2'-OMe modified sugars and one or more 2'-MOE modified sugars. In some embodiments, the last several sugars comprise one or more 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 aliphatic), or bicyclic sugars as described herein (e.g., LNA, cEt, etc.). In some embodiments, the last several sugars include one or more 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the last several sugars comprise one or more 2'-OMe modified sugars. In some embodiments, the last several sugars comprise one or more 2'-MOE modified sugars. In some embodiments, the last several sugars comprise one or more 2'-OMe modified sugars and one or more 2'-MOE modified sugars. In some embodiments, the last several sugars are independently 2'-OMe modified sugars. In some embodiments, the first several sugars comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.) contiguous bicyclic sugars or 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the first several sugars include two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.) consecutive 2'-OR modified sugars (wherein R is an optionally substituted C 1~6In some embodiments, the first several sugars comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.) contiguous 2'-OMe modified sugars, where each modified sugar is independently a 2'-OMe modified sugar or a 2'-MOE modified sugar. In some embodiments, the first several sugars comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.) contiguous 2'-OMe modified sugars. In some embodiments, the first several sugars comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.) contiguous 2'-MOE modified sugars. In some embodiments, the last several sugars comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.) contiguous 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the last several sugars comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.) contiguous modified sugars, each modified sugar being independently a 2'-OMe modified sugar or a 2'-MOE modified sugar. In some embodiments, the last several sugars comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.) contiguous 2'-OMe modified sugars. In some embodiments, the last several sugars comprise three or more contiguous 2'-OMe modified sugars. In some embodiments, the last several sugars comprise four or more contiguous 2'-OMe modified sugars. In some embodiments, the last several sugars comprise five or more contiguous 2'-OMe modified sugars. In some embodiments, the last several sugars comprise six or more contiguous 2'-OMe modified sugars. In some embodiments, the last several sugars include two or more (eg, 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.) consecutive 2'-MOE modified sugars.

[0144] In some embodiments, one or more (1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of the first several (1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) sugars are modified sugars. In some embodiments, one or more (1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of the first several (1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) sugars are 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 aliphatic), and bicyclic sugars (e.g., 2'-O-CH 2 -4'(in the formula, -CH 2 - is a modified sugar independently selected from a sugar (e.g., an LNA sugar, a cET sugar (e.g., (S)-cET)) that includes an optionally substituted C 1~6 In some embodiments, three or more of the first several sugars are 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, four or more of the first several sugars are 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, one or more sugars are contiguous. In some embodiments, the first 1, 2, 3, or 4 sugars are modified sugars. In some embodiments, the first two sugars are 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the first three sugars are modified sugars independently selected from 2'-OR modified sugars (wherein R is an optionally substituted C 1~6In some embodiments, the first four sugars are modified sugars independently selected from 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, each 2'-OR modified sugar is independently a 2'-OMe or a 2'-MOE modified sugar. In some embodiments, each bicyclic sugar is independently an LNA sugar or a cEt sugar. In some embodiments, one or more (1, 2, 3, 4, or 5) sugars of the first several sugars, or each of the first several (e.g., 1, 2, 3, 4, or 5) sugars, is independently a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6aliphatic). In some embodiments, one or more (1, 2, 3, 4, or 5) sugars of the first several sugars or each of the first several (e.g., 1, 2, 3, 4, or 5) sugars are independently 2'-OMe or 2'-MOE modified sugars. In some embodiments, one or more (1, 2, 3, 4, or 5) sugars of the first several sugars or each of the first several (e.g., 1, 2, 3, 4, or 5) sugars are independently 2'-OMe modified sugars. In some embodiments, one or more (1, 2, 3, 4, or 5) sugars of the first several sugars or each of the first several (e.g., 1, 2, 3, 4, or 5) sugars are independently 2'-OMe modified sugars. In some embodiments, one or more (1, 2, 3, 4, or 5) sugars of the first several sugars or each of the first several (e.g., 1, 2, 3, 4, or 5) sugars are independently 2'-MOE modified sugars. In some embodiments, the first 1, 2, 3, 4, or more sugars are independently 2'-OMe modified sugars. In some embodiments, the first sugar is a 2'-OMe modified sugar. In some embodiments, the first two sugars are independently 2'-OMe modified sugars. In some embodiments, the first three sugars are independently 2'-OMe modified sugars. In some embodiments, the first four sugars are independently 2'-OMe modified sugars. In some embodiments, the first 1, 2, 3, 4, or more sugars are independently 2'-MOE modified sugars. In some embodiments, the first sugar is a 2'-MOE modified sugar. In some embodiments, the first two sugars are independently 2'-MOE modified sugars. In some embodiments, the first three sugars are independently 2'-MOE modified sugars. In some embodiments, the first four sugars are independently 2'-MOE modified sugars. In some embodiments, each such modified sugar is independently a nucleoside sugar in which the nucleobase is an optionally substituted or protected A, T, C, G, or U, or an optionally substituted or protected tautomer of A, T, C, G, or U. In some embodiments, one or more such sugars are independently attached to a PN bond. In some embodiments, one or more such sugars are independently attached to a non-negatively charged internucleotide linkage.In some embodiments, one or more such sugars are independently linked to a neutral internucleotide linkage, such as n001. In some embodiments, a non-negatively charged internucleotide linkage or a neutral internucleotide linkage, such as n001, is chiral controlled. In some embodiments, this is Rp. In some embodiments, one or more such sugars are independently linked to a PS linkage. In some embodiments, one or more such sugars are independently linked to a phosphorothioate internucleotide linkage. In some embodiments, a PS linkage, e.g., a phosphorothioate internucleotide linkage, is chiral controlled. In some embodiments, this is Sp. In some embodiments, the internucleotide linkage between the first nucleoside and the second nucleoside is a non-negatively charged internucleotide linkage, as described herein. In some embodiments, this is a neutral internucleotide linkage. In some embodiments, this is a PN linkage. In some embodiments, this is a phosphorylguanidine internucleotide linkage. In some embodiments, this is n001. In some embodiments, this is chiral controlled. In some embodiments, this is Rp. In some embodiments, one or more of the first several, or each internucleotide linkage attached to the first several modified sugar-containing nucleosides, except for the internucleotide linkage between the first and second nucleosides, is independently a PS linkage, e.g., a phosphorothioate internucleotide linkage. In some embodiments, each is chiral controlled. In some embodiments, each is Sp. In some embodiments, the first nucleoside is linked to an additional moiety (e.g., Mod001) through its 5' terminal carbon (in some embodiments, through a phosphate group), optionally through a linker (e.g., L001). In some embodiments, the first several are the first 3, 4, 5, 6, etc.

[0145] In some embodiments, one or more (1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of the last several (1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) sugars are modified sugars. In some embodiments, one or more (1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of the last several (1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) sugars are 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 aliphatic), and bicyclic sugars (e.g., 2'-O-CH 2 -4'(in the formula, -CH 2 - is a modified sugar independently selected from a sugar (e.g., an LNA sugar, a cET sugar (e.g., (S)-cET)) that includes an optionally substituted C 1~6 In some embodiments, three or more of the last several sugars are 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, four or more of the last several sugars are 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, one or more sugars are contiguous. In some embodiments, the last 1, 2, 3, or 4 sugars are modified sugars. In some embodiments, the last two sugars are 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the last three sugars are modified sugars independently selected from 2'-OR modified sugars (wherein R is an optionally substituted C 1~6In some embodiments, the last four sugars are modified sugars independently selected from 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, each 2'-OR modified sugar is independently a 2'-OMe or a 2'-MOE modified sugar. In some embodiments, each bicyclic sugar is independently an LNA sugar or a cEt sugar. In some embodiments, one or more (1, 2, 3, 4, or 5) of the last several sugars, or each of the last several (e.g., 1, 2, 3, 4, or 5) sugars, is independently a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6aliphatic). In some embodiments, one or more (1, 2, 3, 4, or 5) sugars of the last several sugars or each of the last several (e.g., 1, 2, 3, 4, or 5) sugars are independently 2'-OMe or 2'-MOE modified sugars. ... modified sugars. In some embodiments, one or more (1, 2, 3, 4, or 5) sugars of the last several sugars or each of the last several (e.g., 1, 2, 3, 4, or 5) sugars are independently 2'-MOE modified sugars. In some embodiments, the last 1, 2, 3, 4, or more sugars are independently 2'-OMe modified sugars. In some embodiments, the last sugar is a 2'-OMe modified sugar. In some embodiments, the last two sugars are independently, 2'-OMe modified sugars. In some embodiments, the last three sugars are independently, 2'-OMe modified sugars. In some embodiments, the last four sugars are independently, 2'-OMe modified sugars. In some embodiments, the last 1, 2, 3, 4, or more sugars are independently, 2'-MOE modified sugars. In some embodiments, the last sugar is a 2'-MOE modified sugar. In some embodiments, the last two sugars are independently, 2'-MOE modified sugars. In some embodiments, the last three sugars are independently, 2'-MOE modified sugars. In some embodiments, the last four sugars are independently, 2'-MOE modified sugars. In some embodiments, each such modified sugar is independently a nucleoside sugar in which the nucleobase is an optionally substituted or protected A, T, C, G, or U, or an optionally substituted or protected tautomer of A, T, C, G, or U. In some embodiments, one or more such sugars are independently attached to a PN bond. In some embodiments, one or more such sugars are independently attached to a non-negatively charged internucleotide linkage.In some embodiments, one or more such sugars are independently linked to a neutral internucleotide linkage, such as n001. In some embodiments, a non-negatively charged internucleotide linkage or a neutral internucleotide linkage, such as n001, is chiral controlled. In some embodiments, it is Rp. In some embodiments, one or more such sugars are independently linked to a PS linkage. In some embodiments, one or more such sugars are independently linked to a phosphorothioate internucleotide linkage. In some embodiments, the PS linkage is chiral controlled. In some embodiments, the phosphorothioate internucleotide linkage is chiral controlled. In some embodiments, it is Sp. In some embodiments, the internucleotide linkage between the last nucleoside and the penultimate nucleoside is a non-negatively charged internucleotide linkage, as described herein. In some embodiments, it is a neutral internucleotide linkage. In some embodiments, it is a PN linkage. In some embodiments, it is a phosphorylguanidine internucleotide linkage. In some embodiments, this sugar is n001. In some embodiments, it is chiral controlled. In some embodiments, it is Rp. In some embodiments, each internucleotide linkage attached to one or more of the last several or the last several nucleosides containing modified sugars, except for the internucleotide linkage between the last and penultimate nucleosides, is independently a phosphorothioate internucleotide linkage. In some embodiments, each is chiral controlled. In some embodiments, each is Sp. In some embodiments, the last several are the last 3, 4, 5, etc.

[0146] In some embodiments, the sugar at the +1 position is a 2'-F modified sugar. In some embodiments, the sugar at the +1 position is a natural DNA sugar. In some embodiments, the sugar at the 0 position is a natural DNA sugar (the nucleoside at the 0 position is opposite the target adenosine when aligned). In some embodiments, the sugar at the -1 position is a DNA sugar. In some embodiments, the sugar at the -2 position is a 2'-OR modified sugar (where R is an optionally substituted C 1~6 aliphatic), or bicyclic sugars (e.g., 2'-O-CH 2 -4'(in the formula, -CH 2 - is an optionally substituted sugar (e.g., an LNA sugar, a cET sugar (e.g., (S)-cEt)). In some embodiments, this is a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, it is a 2'-OMe modified sugar. In some embodiments, it is a 2'-MOE modified sugar. In some embodiments, it is a bicyclic sugar. In some embodiments, it is an LNA sugar. In some embodiments, it is a cEt sugar. In some embodiments, the sugar at the -3 position is a 2'-F modified sugar. In some embodiments, each sugar after the -3 position (e.g., at the -4, -5, -6 positions, etc.) is independently a 2'-OR modified sugar (where R is an optionally substituted C 1~6 aliphatic), or bicyclic sugars (e.g., 2'-O-CH 2 -4'(in the formula, -CH 2 - is an optionally substituted sugar (e.g., an LNA sugar, a cET sugar (e.g., (S)-cEt)). In some embodiments, each is independently a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6aliphatic), or bicyclic sugar. In some embodiments, each is independently a 2'-OMe or 2'-MOE modified sugar. In some embodiments, each is a 2'-OMe modified sugar. In some embodiments, each is a 2'-MOE modified sugar. In some embodiments, one or more are independently a 2'-OMe modified sugar and one or more are independently a 2'-MOE modified sugar. In some embodiments, as described herein, the internucleotide linkage between the nucleoside at the -1 position and the nucleoside at the -2 position is a non-negatively charged internucleotide linkage. In some embodiments, it is a neutral internucleotide linkage. In some embodiments, it is a phosphorylguanidine internucleotide linkage. In some embodiments, it is n001. In some embodiments, it is chiral controlled. In some embodiments, it is Sp. In some embodiments, it is Rp. In some embodiments, the internucleotide bond between the nucleoside at the -2 position and the nucleoside at the -3 position is a natural phosphate bond. In some embodiments, the internucleotide bond between the last nucleoside and the penultimate nucleoside is a non-negatively charged internucleotide bond as described herein. In some embodiments, it is a neutral internucleotide bond. In some embodiments, it is a phosphorylguanidine internucleotide bond. In some embodiments, it is n001. In some embodiments, it is chiral controlled. In some embodiments, it is Rp. In some embodiments, each internucleotide bond between nucleosides to the 3' side of the nucleoside opposite the target adenosine is independently a phosphorothioate internucleotide bond, except for those between the nucleoside at the -1 position and the nucleoside at the -2 position, between the nucleoside at the -2 position and the nucleoside at the -3 position, and between the last nucleoside and the penultimate nucleoside. In some embodiments, each phosphorothioate internucleotide linkage is chiral controlled. In some embodiments, each is Sp.

[0147] In some embodiments, the first and / or last one or some of the sugars are modified sugars, e.g., bicyclic sugars and / or 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 aliphatic) (e.g., 2'-OMe modified sugars, 2'-MOE modified sugars, etc.). In some embodiments, such sugars may increase the stability, affinity, and / or activity of the oligonucleotide. In some embodiments, when conjugated to one or more additional chemical moieties, the sugars at the 5' and / or 3' ends of the oligonucleotide are bicyclic sugars or 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the 5'-terminal sugar is not a bicyclic sugar or a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, such 5' terminal sugar is not linked to an additional chemical moiety. In some embodiments, the 5' terminal sugar is a 2'-F modified sugar. In some embodiments, the 5' terminal sugar is a 2'-F modified sugar conjugated to an additional chemical moiety. In some embodiments, the 3' terminal sugar is a bicyclic sugar or a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, such 3' terminal sugar is not linked to an additional chemical moiety. In some embodiments, the 3' terminal sugar is a 2'-F modified sugar. In some embodiments, the 3' terminal sugar is a 2'-F modified sugar conjugated to an additional chemical moiety. In some embodiments, the last several sugars are sugars 3' to the nucleoside opposite the target adenosine (e.g., N -1 , N -2In some embodiments, the last several sugars or the 3' sugars comprise one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more) 2'-F modified sugars. In some embodiments, the last several sugars or the 3' sugars comprise two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more) consecutive 2'-F modified sugars. In some embodiments, the last several sugars or the 3' sugars comprise one or more 2'-F modified sugars or two or more consecutive 2'-F modified sugars, and the last nucleoside sugar of the oligonucleotide is a bicyclic sugar or a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, the 2'-OR modified sugar is a 2'-OMe modified sugar or a 2'-MOE modified sugar, in some embodiments it is a 2'-OMe modified sugar, in some embodiments it is a 2'-MOE modified sugar, as described herein. In some embodiments, the last several sugars, or the sugar on the 3' side, comprise one or more 2'-F modified sugars, or two or more consecutive 2'-F modified sugars, and the sugar of the last nucleoside of the oligonucleotide is a 2'-OR modified sugar, where R is an optionally substituted C 1~6aliphatic). In some embodiments, the last several sugars or the sugar on the 3' side contain one or more 2'-F modified sugars or two or more consecutive 2'-F modified sugars, and the sugar of the last nucleoside of the oligonucleotide is a 2'-OMe modified sugar or a 2'-MOE modified sugar. In some embodiments, the last several sugars or the sugar on the 3' side contain one or more 2'-F modified sugars or two or more consecutive 2'-F modified sugars, and the sugar of the last nucleoside of the oligonucleotide is a 2'-OMe modified sugar. In some embodiments, the last several sugars or the sugar on the 3' side contain one or more 2'-F modified sugars or two or more consecutive 2'-F modified sugars, and the sugar of the last nucleoside of the oligonucleotide is a 2'-MOE modified sugar. In some embodiments, no more than two nucleosides 3' to the nucleoside opposite the adenosine independently have a 2'-F modified sugar. In some embodiments, these are at the -4 and -5 positions. In some embodiments, these are the penultimate and third nucleosides of the oligonucleotide. In some embodiments, no more than one nucleoside 3' to the nucleoside opposite the adenosine has a 2'-F modified sugar. In some embodiments, this is at the -3 position. In some embodiments, this is the fourth penultimate nucleoside of the oligonucleotide.

[0148] In some embodiments, a bicyclic sugar or a 2'-OR modified sugar, where R is an optionally substituted C 1~6aliphatic) are present in a region that includes one or more (e.g., 1-30, 1-25, 1-20, 1-15, 1-10, 2-30, 2-25, 2-20, 2-25, 2-10, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more) sugars that are 2'-F modified. In some embodiments, a majority of the sugars as described herein in such a region are 2'-F modified sugars. In some embodiments, two or more 2'-F modified sugars are contiguous. In some embodiments, the region is a first domain. In some embodiments, a bicyclic sugar is present in such a region. In some embodiments, a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, 2'-OMe modified sugars are present in such regions. In some embodiments, 2'-MOE modified sugars are present in such regions.

[0149] In some embodiments, one or more sugars at positions -5, -4, -3, +1, +2, +4, +5, +6, +7, and +8 (position 0 is the position of the nucleoside opposite the target adenosine; "+" goes from the nucleoside opposite the target adenosine to the 5' end of the oligonucleotide and "-" goes from the nucleoside opposite the target adenosine to the 3' end of the oligonucleotide; e.g., 5'-N 1 N 0 N -1 In -3', N 0 is at position 0 when it is the nucleoside opposite the target adenosine, and N 1 is present at position +1, and N -1is present at the -1 position) are independently 2'-F modified sugars. In some embodiments, the sugar at the +1 position and one or more sugars at the -5, -4, -3, +2, +4, +5, +6, +7, and +8 positions are independently 2'-F modified sugars. In some embodiments, the sugar at the +1 position and one sugar at the -5, -4, -3, +2, +4, +5, +6, +7, and +8 positions are independently 2'-F modified sugars.

[0150] In some embodiments, an oligonucleotide comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more, such as 2-10, 3-10, 2-5, 2-4, 2-3, 3-5, 3-4, etc.) naturally occurring DNA sugars. In some embodiments, one or more naturally occurring DNA sugars are present at an editing region (e.g., at the +1, 0, and / or -1 positions). In some embodiments, a naturally occurring DNA sugar is present within the first several nucleosides of an oligonucleotide (e.g., the first 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides). In some embodiments, the first, second, and / or third nucleoside of an oligonucleotide, independently, has a naturally occurring DNA sugar. In some embodiments, the natural DNA sugar is linked to a modified internucleotide linkage (e.g., a non-negatively charged internucleotide linkage, a neutral internucleotide linkage, a phosphorylguanidine internucleotide linkage, an n001, or a phosphorothioate internucleotide linkage (in various embodiments, Sp)).

[0151] Oligonucleotides may contain various types of internucleotide linkages. In some embodiments, the oligonucleotides contain one or more modified internucleotide linkages. In some embodiments, the modified internucleotide linkage is a chiral internucleotide linkage. In some embodiments, the modified internucleotide linkage is a phosphorothioate internucleotide linkage. In some embodiments, the modified internucleotide linkage is a non-negatively charged internucleotide linkage. In some embodiments, the modified internucleotide linkage is a neutral internucleotide linkage. In some embodiments, the modified internucleotide linkage is a phosphorylguanidine internucleotide linkage. In some embodiments, the modified internucleotide linkage is n001. In some embodiments, the oligonucleotides contain one or more natural phosphate linkages. In some embodiments, the natural phosphate linkage is linked to a nucleoside that includes a modified sugar that may improve stability (e.g., resistance to nucleases). In some embodiments, the natural phosphate linkage is linked to a bicyclic sugar. In some embodiments, the natural phosphate linkage is linked to a 2'-modified sugar. In some embodiments, the natural phosphate linkage is linked to a 2'-OR modified sugar, where R is an optionally substituted C 1~6aliphatic). In some embodiments, the natural phosphate linkage is attached to a 2'-OMe modified sugar. In some embodiments, the natural phosphate linkage is attached to a 2'-MOE modified sugar. In some embodiments, the oligonucleotide comprises a phosphorothioate internucleotide linkage, a non-negatively charged internucleotide linkage, and a natural phosphate linkage. In some embodiments, the oligonucleotide comprises a phosphorothioate internucleotide linkage, a neutral internucleotide linkage, and a natural phosphate linkage. In some embodiments, the oligonucleotide comprises a phosphorothioate internucleotide linkage, a phosphoryl guanidine internucleotide linkage, and a natural phosphate linkage. In some embodiments, the oligonucleotide comprises a phosphorothioate internucleotide linkage, an n001, and a natural phosphate linkage. In some embodiments, each chiral internucleotide linkage is independently chiral controlled. In some embodiments, one or more chiral internucleotide linkages are not chiral controlled. In some embodiments, each phosphorothioate internucleotide linkage is independently chiral controlled. In some embodiments, each chiral internucleotide linkage is independently chiral controlled. In some embodiments, most or each phosphorothioate internucleotide linkage is Sp as described herein. In some embodiments, most or each non-negatively charged internucleotide linkage (e.g., n001) is Rp. In some embodiments, most or each non-negatively charged internucleotide linkage (e.g., n001) is Sp.

[0152] In some embodiments, the oligonucleotide comprises a phosphorothioate internucleotide linkage and a non-negatively charged internucleotide linkage. In some embodiments, the oligonucleotide comprises a phosphorothioate internucleotide linkage and a neutral internucleotide linkage. In some embodiments, the oligonucleotide comprises a phosphorothioate internucleotide linkage and a phosphorylguanidine internucleotide linkage. In some embodiments, the oligonucleotide comprises a phosphorothioate internucleotide linkage and an n001. In some embodiments, each chiral internucleotide linkage is independently chiral controlled. In some embodiments, one or more chiral internucleotide linkages are not chiral controlled. In some embodiments, each phosphorothioate internucleotide linkage is independently chiral controlled. In some embodiments, each chiral internucleotide linkage is independently chiral controlled. In some embodiments, a majority or each phosphorothioate internucleotide linkage is Sp as described herein. In some embodiments, one or more (e.g., 1, 2, 3, 4, or 5) phosphorothioate internucleotide linkages are Rp. In some embodiments, the majority or each non-negatively charged internucleotide linkage (e.g., n001) is Rp. In some embodiments, the majority or each non-negatively charged internucleotide linkage (e.g., n001) is Sp. In some embodiments, the oligonucleotide does not contain a natural phosphate linkage. In some embodiments, each internucleotide linkage is independently a phosphorothioate or a non-negatively charged internucleotide linkage. In some embodiments, each internucleotide linkage is independently a phosphorothioate or a neutrally charged internucleotide linkage. In some embodiments, each internucleotide linkage is independently a phosphorothioate or a phosphorylguanidine internucleotide linkage. In some embodiments, each internucleotide linkage is independently a phosphorothioate or a n001 internucleotide linkage.In some embodiments, the final internucleotide linkage of the oligonucleotide is a non-negatively charged internucleotide linkage, or a neutral internucleotide linkage, or a phosphorylguanidine internucleotide linkage, or an n001.

[0153] In some embodiments, the oligonucleotide of the present disclosure comprises one or more modified nucleobases.Various modifications can be introduced into sugar and / or nucleobase according to the present disclosure.For example, in some embodiments, the modification is the modification described in US Patent No. 9006198. In some embodiments, modifications can be made to the nucleotides or nucleotides of interest, as described in U.S. Patent Nos. 9,394,333, 9,744,183, 9,605,019, 9,982,257, U.S. Patent Application Publication Nos. 20170037399, 20180216108, 20180216107, U.S. Patent No. 9,598,458, WO 2017 / 062862, WO 2018 / 067973, WO 2017 / 160741, WO 2017 / 192679, WO 2017 / 210647, WO 2018 / 098264, WO 2018 / 2018 / 10771 ... No. 022473, No. 2018 / 223056, No. 2018 / 223073, No. 2018 / 223081, No. 2018 / 237194 , 2019 / 032607, 2019 / 032612, 2019 / 055951, 2019 / 075357, 2019 / The modifications are those described in PCT / US2023 / 018742, PCT / US200185, PCT / US2019 / 217784, PCT / US2019 / 032612, PCT / US2020 / 191252, PCT / US2021 / 071858, PCT / US2021 / 237223, PCT / US2022 / 099159, and / or PCT / US2023 / 018742.

[0154] In some embodiments, the nucleobase in the nucleoside is or comprises a BA ring having the structure BA-I, BA-Ia, BA-Ib, BA-Ic, BA-Id, BA-II, BA-II-a, BA-II-b, BA-II-c, BA-II-d, BA-III, BA-III-a, BA-III-b, BA-III-c, BA-III-d, BA-III-e, BA-IV, BA-IV-a, BA-IV-b, BA-V, BA-Va, BA-Vb, or BA-VI, or a tautomer of the BA ring, wherein the nucleobase is optionally substituted or protected.

[0155] In some embodiments, the sugar is a modified sugar including a 2'-modification, e.g., 2'-F, 2'-OR (where R is an optionally substituted aliphatic), or a bicyclic sugar (e.g., an LNA sugar), or an acyclic sugar (e.g., a UNA sugar).

[0156] In some embodiments, the oligonucleotides provided comprise one or more domains, each of which independently has a certain length, modification, stereochemistry of the phosphorus linkage, etc., as described herein. In some embodiments, the disclosure provides oligonucleotides comprising one or more modified sugars and / or one or more modified internucleotide linkages, the oligonucleotides comprising a first domain and a second domain, each independently comprising one or more nucleobases. In some embodiments, the disclosure provides oligonucleotides comprising one or more domains and / or subdomains as described herein. In some embodiments, the disclosure provides oligonucleotides comprising a first domain as described herein. In some embodiments, the disclosure provides oligonucleotides comprising a second domain as described herein. In some embodiments, the disclosure provides oligonucleotides comprising a first subdomain as described herein. In some embodiments, the disclosure provides oligonucleotides comprising a second subdomain as described herein. In some embodiments, the disclosure provides oligonucleotides comprising a third subdomain as described herein. In some embodiments, the present disclosure provides an oligonucleotide comprising one or more regions independently selected from a first domain, a second domain, a first subdomain, a second subdomain, and a third subdomain, each independently as described herein. In some embodiments, the first domain is linked to the second domain via a linker. In some embodiments, the first domain is directly linked to the linker.

[0157] In some embodiments, an oligonucleotide or portion thereof (e.g., the first domain, the second domain, the first subdomain, the second subdomain, the third subdomain, etc.) comprises a certain level of modified sugars. In some embodiments, the modified sugar comprises a 2'-modification. In some embodiments, the modified sugar is a bicyclic sugar. In some embodiments, the modified sugar is an acyclic sugar (e.g., by cleaving the C2-C3 bond of the corresponding cyclic sugar). In some embodiments, the modified sugar comprises a 5'-modification. Typically, oligonucleotides of the present disclosure have a free 5'-OH at their 5'-terminus and a free 3'-OH at their 3'-terminus, unless otherwise specified, e.g., by context. In some embodiments, the 5' terminal sugar of the oligonucleotide may comprise a modified 5'-OH.

[0158] In some embodiments, the levels are about, e.g., about 5%-100%, about 10%-100%, 20-100%, 30%-100%, 40%-100%, 50%-80%, 50%-85%, 50%-90%, 50%-95%, 60%-80%, 60%-85%, 60%-90%, 60%-95%, 60%-100%, 65%-80%, 65%-85%, 65%-90%, 65%-95%, 65%-100%, 70%-80%, 70%-80%, 70%-80%, 70%-80%, 70%-80%, 70%-80%, 70%-80%, 70%-9 ... %, 70%-85%, 70%-90%, 70%-95%, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the percentage is at least about 50%. In some embodiments, the percentage is at least about 55%. In some embodiments, the percentage is at least about 60%. In some embodiments, the percentage is at least about 65%. In some embodiments, the percentage is at least about 70%. In some embodiments, the percentage is at least about 75%. In some embodiments, the percentage is at least about 80%. In some embodiments, the percentage is at least about 85%. In some embodiments, the percentage is at least about 90%. In some embodiments, the percentage is at least about 95%. In some embodiments, the percentage is about 100%.

[0159] In some embodiments, a majority is at least 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more. In some embodiments, a majority is about 50%-100%, 50%-80%, 50%-85%, 50%-90%, 50%-95%, 60%-80%, 60%-85%, 60%-90%, 60%-95%, 60%-100%, 65%-80%, 65%-85%, 65%-90%, 65%-95%, 65%-100%, 70%-80%, 70%-85%, 70%-90%, 70%-95%, or 70%-100%. , 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, majority is about or at least about 50%. In some embodiments, majority is about or at least about 55%. In some embodiments, majority is about or at least about 60%. In some embodiments, majority is about or at least about 65%. In some embodiments, a majority is about or at least about 70%. In some embodiments, a majority is about or at least about 75%. In some embodiments, a majority is about or at least about 80%. In some embodiments, a majority is about or at least about 85%. In some embodiments, a majority is about or at least about 90%. In some embodiments, a majority is about or at least about 95%.

[0160] In some embodiments, the oligonucleotide or a portion thereof (e.g., the first domain, the second domain, the first subdomain, the second subdomain, the third subdomain, etc.) comprises a particular level of modified internucleotide linkages. In some embodiments, the oligonucleotide or a portion thereof (e.g., the first domain, the second domain, the first subdomain, the second subdomain, the third subdomain, etc.) comprises a particular level of chiral internucleotide linkages. In some embodiments, the levels are about, e.g., about 5%-100%, about 10%-100%, 20-100%, 30%-100%, 40%-100%, 50%-80%, 50%-85%, 50%-90%, 50%-95%, 60%-80%, 60%-85%, 60%-90%, 60%-95%, 60%-100%, 65%-80%, 65%-85%, 65%-90%, 65%-95%, 65%-100%, 70%-80%, 70%-90%, 70%-100%, 80%-80%, 85%-85%, 85%-90%, 85%-90%, 85%-95%, 85%-100%, 80%-100%, 85 ... % to 80%, 70% to 85%, 70% to 90%, 70% to 95%, 70% to 100%, 75% to 80%, 75% to 85%, 75% to 90%, 75% to 95%, 75% to 100%, 80% to 85%, 80% to 90%, 80% to 95%, 80% to 100%, 85% to 90%, 85% to 95%, 85% to 100%, 90% to 95%, 90% to 100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, etc. In some embodiments, the percentage is at least about 50%. In some embodiments, the percentage is at least about 55%. In some embodiments, the percentage is at least about 60%. In some embodiments, the percentage is at least about 65%. In some embodiments, the percentage is at least about 70%. In some embodiments, the percentage is at least about 75%. In some embodiments, the percentage is at least about 80%. In some embodiments, the percentage is at least about 85%. In some embodiments, the percentage is at least about 90%. In some embodiments, the percentage is at least about 95%.In some embodiments, the percentage is about 100%.

[0161] In some embodiments, the oligonucleotide or a portion thereof (e.g., the first domain, the second domain, the first subdomain, the second subdomain, the third subdomain, etc.) comprises a certain level of chiral controlled internucleotide linkages. In some embodiments, the oligonucleotide or a portion thereof (e.g., the first domain, the second domain, the first subdomain, the second subdomain, the third subdomain, etc.) comprises a certain level of Sp internucleotide linkages. In some embodiments, the levels are about, e.g., about 5%-100%, about 10%-100%, 20-100%, 30%-100%, 40%-100%, 50%-80%, 50%-85%, 50%-90%, 50%-95%, 60%-80%, 60%-85%, 60%-90%, 60%-95%, 60%-100%, 65%-80%, 65%-85%, 65%-90%, 65%-95%, 65%-100%, 70%-80%, 70%-90%, 70%-100%, 80%-80%, 85%-85%, 85%-90%, 85%-90%, 85%-95%, 85%-100%, 80%-100%, 85 ... %~80%, 70%~85%, 70%~90%, 70%~95%, 70%~100%, 75%~80%, 75%~85%, 75%~90%, 75%~95%, 75%~100%, 80%~85%, 80%~90%, 80%~95%, 80%~100%, 85%~90%, 85%~95%, 85%~100%, 90%~95%, 90%~100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% etc.In some embodiments, the level is about, e.g., about 5% to 100%, about 10% to 100%, 20 to 100%, 30% to 100%, 40% to 100%, 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65% to 100%, 70%-80%, 70%-85%, 70%-90%, 70%-95%, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, etc. In some embodiments, the percentage is at least about 50%. In some embodiments, the percentage is at least about 55%. In some embodiments, the percentage is at least about 60%. In some embodiments, the percentage is at least about 65%. In some embodiments, the percentage is at least about 70%. In some embodiments, the percentage is at least about 75%. In some embodiments, the percentage is at least about 80%. In some embodiments, the percentage is at least about 85%. In some embodiments, the percentage is at least about 90%. In some embodiments, the percentage is at least about 95%. In some embodiments, the percentage is about 100%.

[0162] In some embodiments, the oligonucleotide or a portion thereof (e.g., the first domain, the second domain, the first subdomain, the second subdomain, the third subdomain, etc.) comprises a certain level of Sp internucleotide linkages. In some embodiments, the level may be about, e.g., about 5%-100%, about 10%-100%, 20%-100%, 30%-100%, 40%-100%, 50%-80%, 50%-85%, 50%-90%, 50%-95%, 60%-80%, 60%-85%, 60%-90%, 60%-95%, 60%-100%, 65%-80%, 65%-85%, 65%-90%, 65%-95%, 65%-100%, 70%-80%, 75%-90%, 75%-100%, 80%-85%, 80%-90%, 80%-10 ... %~80%, 70%~85%, 70%~90%, 70%~95%, 70%~100%, 75%~80%, 75%~85%, 75%~90%, 75%~95%, 75%~100%, 80%~85%, 80%~90%, 80%~95%, 80%~100%, 85%~90%, 85%~95%, 85%~100%, 90%~95%, 90%~100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% etc. In some embodiments, the level is about, e.g., about 5% to 100%, about 10% to 100%, 20 to 100%, 30% to 100%, 40% to 100%, 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65% to 100%, 70%~80%, 70%~85%, 70%~90%, 70%~95%, 70%~100%, 75%~80%, 75%~85%, 75%~90%, 75%~95%, 75%~100%, 80%~85%, 80%~90%, 80%~95%, 80%~100%, 85%~90%, 85%~95%, 85%~100%, 90%~95%, 90%~100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.In some embodiments, the level is about, e.g., about 5% to 100%, about 10% to 100%, 20 to 100%, 30% to 100%, 40% to 100%, 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65% to 100%, respectively, of all chiral controlled internucleotide linkages in the oligonucleotide or portion thereof. %, 70%-80%, 70%-85%, 70%-90%, 70%-95%, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the percentage is at least about 50%. In some embodiments, the percentage is at least about 55%. In some embodiments, the percentage is at least about 60%. In some embodiments, the percentage is at least about 65%. In some embodiments, the percentage is at least about 70%. In some embodiments, the percentage is at least about 75%. In some embodiments, the percentage is at least about 80%. In some embodiments, the percentage is at least about 85%. In some embodiments, the percentage is at least about 90%. In some embodiments, the percentage is at least about 95%. In some embodiments, the percentage is about 100%. In some embodiments, about 1 to 50, 1 to 40, 1 to 30, e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, internucleotide linkages are independently Sp chiral internucleotide linkages.In many embodiments, a high percentage of Sp internucleotide linkages (e.g., compared to Rp internucleotide linkages and / or natural phosphate linkages) in an oligonucleotide or certain portions thereof can result in improved properties and / or activity (e.g., increased stability and / or increased adenosine editing activity).

[0163] In some embodiments, the oligonucleotide or a portion thereof (e.g., the first domain, the second domain, the first subdomain, the second subdomain, the third subdomain, etc.) comprises a certain level of Rp internucleotide linkages. In some embodiments, the level may be about, e.g., about 5%-100%, about 10%-100%, 20%-100%, 30%-100%, 40%-100%, 50%-80%, 50%-85%, 50%-90%, 50%-95%, 60%-80%, 60%-85%, 60%-90%, 60%-95%, 60%-100%, 65%-80%, 65%-85%, 65%-90%, 65%-95%, 65%-100%, 70%-80%, 75%-90%, 75%-100%, 80%-80%, 85%-90%, 85%-100%, 85%-100%, 90 ... %~80%, 70%~85%, 70%~90%, 70%~95%, 70%~100%, 75%~80%, 75%~85%, 75%~90%, 75%~95%, 75%~100%, 80%~85%, 80%~90%, 80%~95%, 80%~100%, 85%~90%, 85%~95%, 85%~100%, 90%~95%, 90%~100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% etc. In some embodiments, the level is about, e.g., about 5% to 100%, about 10% to 100%, 20 to 100%, 30% to 100%, 40% to 100%, 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65% to 100%, 70%~80%, 70%~85%, 70%~90%, 70%~95%, 70%~100%, 75%~80%, 75%~85%, 75%~90%, 75%~95%, 75%~100%, 80%~85%, 80%~90%, 80%~95%, 80%~100%, 85%~90%, 85%~95%, 85%~100%, 90%~95%, 90%~100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.In some embodiments, the level is about, e.g., about 5% to 100%, about 10% to 100%, 20 to 100%, 30% to 100%, 40% to 100%, 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65% to 100%, respectively, of all chiral controlled internucleotide linkages in the oligonucleotide or portion thereof. %, 70%-80%, 70%-85%, 70%-90%, 70%-95%, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the percentage is at least about 50%. In some embodiments, the percentage is at least about 55%. In some embodiments, the percentage is at least about 60%. In some embodiments, the percentage is at least about 65%. In some embodiments, the percentage is at least about 70%. In some embodiments, the percentage is at least about 75%. In some embodiments, the percentage is at least about 80%. In some embodiments, the percentage is at least about 85%. In some embodiments, the percentage is at least about 90%. In some embodiments, the percentage is at least about 95%. In some embodiments, the percentage is about 100%. In some embodiments, the percentage is about 5% or less than about 5%. In some embodiments, the percentage is about 10% or less than about 10%. In some embodiments, the percentage is about 15% or less than about 15%. In some embodiments, the percentage is about 20% or less than about 20%. In some embodiments, the percentage is about 25% or less than about 25%.In some embodiments, the percentage is about 30% or less than about 30%. In some embodiments, the percentage is about 35% or less than about 35%. In some embodiments, the percentage is about 40% or less than about 40%. In some embodiments, the percentage is about 45% or less than about 45%. In some embodiments, the percentage is about 50% or less than about 50%. In some embodiments, about 1-50, 1-40, 1-30, e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 internucleotide linkages are independently Rp chiral internucleotide linkages. In some embodiments, this number is about 1 or less than about 1. In some embodiments, this number is about 2 or less than about 2. In some embodiments, this number is about 3 or less than about 3. In some embodiments, this number is about 4 or not more than about 4. In some embodiments, this number is about 5 or not more than about 5. In some embodiments, this number is about 6 or not more than about 6. In some embodiments, this number is about 7 or not more than about 7. In some embodiments, this number is about 8 or not more than about 8. In some embodiments, this number is about 9 or not more than about 9. In some embodiments, this number is about 10 or not more than about 10.

[0164] Without wishing to be bound by a particular theory, in some examples, the Rp and Sp configurations of the internucleotide bond may affect the structural changes of the helical structure of the double-stranded complex formed by the oligonucleotide and the target nucleic acid such as RNA, and the ADAR protein may recognize and interact with various targets (e.g., double-stranded complex formed by the oligonucleotide and the target nucleic acid such as RNA) through multiple domains. In some embodiments, the oligonucleotides and compositions thereof provided facilitate and / or enhance the interaction profile of the oligonucleotide, the target nucleic acid, and / or the ADAR protein, resulting in efficient adenosine modification by the ADAR protein through the incorporation of various modifications and / or stereochemical control.

[0165] In some embodiments, an oligonucleotide may have or include a base sequence; an internucleotide linkage, a base modification, a sugar modification, an additional chemical moiety, or a pattern thereof; and / or any other structural element described herein (e.g., in the Tables).

[0166] In some embodiments, a provided oligonucleotide or composition is characterized in that when it is contacted in a system (e.g., an ADAR-mediated deamination system) with a target nucleic acid that contains a target adenosine, modification of the target adenosine (e.g., deamination of target A) is improved compared to that observed under reference conditions (e.g., selected from the group consisting of the absence of the composition, the presence of a reference oligonucleotide or composition, and combinations thereof). In some embodiments, the modification (e.g., ADAR-mediated deamination (e.g., endogenous ADAR-mediated deamination)) is increased by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000 fold or more.

[0167] In some embodiments, the oligonucleotide is provided as a salt form. In some embodiments, the oligonucleotide is provided as a salt that includes a negatively charged internucleotide bond (e.g., a phosphorothioate internucleotide bond, a natural phosphate bond, etc.) that is present in a salt form. In some embodiments, the oligonucleotide is provided as a pharma- ceutically acceptable salt. In some embodiments, the oligonucleotide is provided as a metal salt. In some embodiments, the oligonucleotide is provided as a sodium salt. In some embodiments, the oligonucleotide is provided as an ammonium salt. In some embodiments, the oligonucleotide is provided as a metal salt (e.g., a sodium salt), and each negatively charged internucleotide bond is independently present in a salt form (e.g., -OP(O)(SNa)-O- for a phosphorothioate internucleotide bond, -OP(O)(ONa)-O- for a natural phosphate bond, etc., for a sodium salt).

[0168] In some embodiments, the oligonucleotide is chiral controlled and contains one or more chiral controlled internucleotide linkages. In some embodiments, the oligonucleotide provided is stereochemically pure. In some embodiments, the oligonucleotide or composition provided is stereochemically pure from other stereoisomers. In some embodiments, the present disclosure provides chiral controlled oligonucleotide compositions.

[0169] As described herein, oligonucleotides of the disclosure may be provided with high purity (e.g., 50%-100%). In some embodiments, oligonucleotides of the disclosure are of high stereochemical purity (e.g., 50%-100%). In some embodiments, the oligonucleotides in the provided compositions are of high stereochemical purity (e.g., a high percentage (e.g., 50%-100%) of a stereoisomer compared to other stereoisomers of the same oligonucleotide). In some embodiments, the percentage is at least or about 50%. In some embodiments, the percentage is at least or about 60%. In some embodiments, the percentage is at least or about 70%. In some embodiments, the percentage is at least or about 75%. In some embodiments, the percentage is at least or about 80%. In some embodiments, the percentage is at least or about 85%. In some embodiments, the percentage is at least or about 90%. In some embodiments, the percentage is at least or about 95%.

[0170] First Domain As described herein, in some embodiments, the oligonucleotide comprises a first domain and a second domain. In some embodiments, the oligonucleotide consists of a first domain and a second domain. By way of example, certain embodiments are described below.

[0171] In some embodiments, the length of the first domain is about or at least about 5-100, 5-50, 5-40, 5-30, 5-20, 10-100, 10-50, 10-40, 10-30, 10-20, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases. In some embodiments, the first domain has a length of about 2 to 50, e.g., about 5, 6, 7, 8, 9, or 10 to about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50 or more nucleobases. In some embodiments, the first domain has a length of about 5 to 30 nucleobases. In some embodiments, the first domain has a length of about 10 to 30 nucleobases. In some embodiments, the first domain has a length of about 5 to 20 nucleobases. In some embodiments, the first domain has a length of about 7-16 nucleobases. In some embodiments, the first domain has a length of 7 nucleobases. In some embodiments, the first domain has a length of 8 nucleobases. In some embodiments, the first domain has a length of 9 nucleobases. In some embodiments, the first domain has a length of 10 nucleobases. In some embodiments, the first domain has a length of 11 nucleobases. In some embodiments, the first domain has a length of 12 nucleobases. In some embodiments, the first domain has a length of 13 nucleobases. In some embodiments, the first domain has a length of 14 nucleobases. In some embodiments, the first domain has a length of 15 nucleobases. In some embodiments, the first domain has a length of 16 nucleobases. In some embodiments, the first domain has a length of 17 nucleobases. In some embodiments, the first domain has a length of 18 nucleobases.In some embodiments, the first domain has a length of 19 nucleobases. In some embodiments, the first domain has a length of 20 nucleobases. In some embodiments, the first domain has a length of 21 or more nucleobases. In some embodiments, the first domain has the same length as the first portion.

[0172] In some embodiments, about or at least about 5-100, 5-50, 5-40, 5-30, 5-20, 10-100, 10-50, 10-40, 10-30, 10-20, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases of the first domain are each independently an optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, each nucleobase of the first domain is independently an optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, about 10-30 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, about 10-25 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, about 10 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, about 11 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, about 12 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or optionally substituted tautomers of A, T, C, G, or U. In some embodiments, about 13 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or optionally substituted tautomers of A, T, C, G, or U.In some embodiments, about 14 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, about 15 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, about 16 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, about 17 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, about 18 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, about 19 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, about 20 nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, about 21 or more nucleobases of the first domain are each independently optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, each such nucleobase is independently an optionally substituted A, T, C, G, or U. In some embodiments, each such nucleobase is independently A, T, C, G, or U.

[0173] The base sequence of the first domain typically has sufficient complementarity to the first portion so that the first domain can form a duplex with the first portion of the target nucleic acid. In some embodiments, the base sequence of the first domain is about 70% to 100%, for example, about or at least about 70%, 75%, 80%, 85%, 90%, 95%, or about 100% complementary to the base sequence of the first portion. In some embodiments, the complementarity is about 70% or more. In some embodiments, the complementarity is about 75% or more. In some embodiments, the complementarity is about 80% or more. In some embodiments, the complementarity is about 85% or more. In some embodiments, the complementarity is about 90% or more. In some embodiments, the complementarity is about 95% or more. In some embodiments, the complementarity is about 100%. In some embodiments, there are one or more mismatches. In some embodiments, there are one or more wobbles. In some embodiments, there are one or more bulges. In some embodiments, the first nucleobase from the 5' end of the first domain is complementary to the nucleobase of the first portion. In some embodiments, the first nucleobase from the 3' end of the first domain is complementary to the nucleobase of the first portion. In some embodiments, when the oligonucleotide is administered or delivered to a system that includes a target nucleic acid or a fragment thereof that includes the first portion, the first domain can selectively form a duplex with the first portion. In some embodiments, when the oligonucleotide is administered or delivered to a system that expresses a target nucleic acid or a fragment thereof that includes the portion, the first domain can selectively form a duplex with the first portion. In some embodiments, the base sequence of the first portion distinguishes the first portion from the remainder of the target nucleic acid. In some embodiments, the base sequence of the first portion distinguishes the first portion from other transcripts in the system. In some embodiments, the base sequence of the first portion distinguishes the first portion from other RNAs in the system. In some embodiments, the base sequence of the first portion distinguishes the first portion from other nucleobases in the system.

[0174] In some embodiments, the first domain is about or at least about 5-95%, 10%-90%, 20%-80%, 30%-70%, 40%-70%, 40%-60%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% of the oligonucleotide. In some embodiments, the percentage is about 30%-80%. In some embodiments, the percentage is about 30%-70%. In some embodiments, the percentage is about 40%-60%. In some embodiments, the percentage is about 20%. In some embodiments, the percentage is about 25%. In some embodiments, the percentage is about 30%. In some embodiments, the percentage is about 35%. In some embodiments, the percentage is about 40%. In some embodiments, the percentage is about 45%. In some embodiments, the percentage is about 50%. In some embodiments, the percentage is about 55%. In some embodiments, the percentage is about 60%. In some embodiments, the percentage is about 65%. In some embodiments, the percentage is about 70%. In some embodiments, the percentage is about 75%. In some embodiments, the percentage is about 80%. In some embodiments, the percentage is about 85%. In some embodiments, the percentage is about 90%.

[0175] In some embodiments, when the oligonucleotide is aligned with a target nucleic acid in terms of complementarity, there are one or more (e.g., 1-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.) mismatches in the first domain. In some embodiments, there is one mismatch. In some embodiments, there are two mismatches. In some embodiments, there are three mismatches. In some embodiments, there are four mismatches. In some embodiments, there are five mismatches. In some embodiments, there are six mismatches. In some embodiments, there are seven mismatches. In some embodiments, there are eight mismatches. In some embodiments, there are nine mismatches. In some embodiments, there are ten mismatches.

[0176] In some embodiments, when the oligonucleotide is aligned with a target nucleic acid in terms of complementarity, there are one or more (e.g., 1-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.) wobbles in the first domain. In some embodiments, there is 1 wobble. In some embodiments, there are 2 wobbles. In some embodiments, there are 3 wobbles. In some embodiments, there are 4 wobbles. In some embodiments, there are 5 wobbles. In some embodiments, there are 6 wobbles. In some embodiments, there are 7 wobbles. In some embodiments, there are 8 wobbles. In some embodiments, there are 9 wobbles. In some embodiments, there are 10 wobbles.

[0177] In some embodiments, the duplex of the oligonucleotide and the target nucleic acid in the first domain region contains one or more bulges, each independently containing one or more mismatches that are not wobble. In some embodiments, there are 0 to 10 bulges (e.g., 0 to 1, 0 to 2, 0 to 3, 0 to 4, 0 to 5, 0 to 6, 0 to 7, 0 to 8, 0 to 9, 0 to 10, 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 2 to 3, 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 2 to 10, 3 to 4, 3 to 5, 3 to 6, 3 to 7, 3 to 8, 3 to 9, 3 to 10, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.). In some embodiments, this number is 0. In some embodiments, this number is 1. In some embodiments, this number is 2. In some embodiments, this number is 3. In some embodiments, this number is 4. In some embodiments, this number is 5.

[0178] In some embodiments, the first domain is perfectly complementary to the target nucleic acid.

[0179] In some embodiments the first domain comprises one or more modified nucleobases.

[0180] In some embodiments, the second domain comprises one or more sugars that comprise two 2'-H (e.g., natural DNA sugars). In some embodiments, the second domain comprises one or more sugars that comprise a 2'-OH (e.g., natural RNA sugars). In some embodiments, the first domain comprises one or more modified sugars. In some embodiments, the modified sugar comprises a 2'-modification. In some embodiments, the modified sugar is a bicyclic sugar (e.g., an LNA sugar). In some embodiments, the modified sugar is an acyclic sugar (e.g., by cleaving the C2-C3 bond of the corresponding cyclic sugar).

[0181] In some embodiments, the first domain comprises about 1 to 50 (e.g., about 5, 6, 7, 8, 9, or 10 to about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, etc., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, etc.) modified sugars. In some embodiments, the first domain comprises about 1 to 50 (e.g., about 5, 6, 7, 8, 9, or 10 to about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, etc., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, etc.) modified sugars having 2'-F modifications.In some embodiments, the first domain comprises about 2 to 50 (e.g., about 2, 3, 4, 5, 6, 7, 8, 9, or 10 to about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, etc., 2-40, 2-30, 2-25, 2-20, 2-15, 2-10, 3-40, 3-30, 3-25, 3-20, 3-15, 3-10, 4-40, 4-3 0, 4~25, 4~20, 4~15, 4~10, 5~40, 5~30, 5~25, 5~20, 5~15, 5~10, 6~40, 6~30, 6~25, 6~20, 6~15, 6~10, 7~40, 7~30, 7~25, 7~20, 7~15, 7~10, 8~40, 8~30, 8~25, 8~20, 8~15, In some embodiments, the first domain comprises about 8-10, 9-40, 9-30, 9-25, 9-20, 9-15, 9-10, 10-40, 10-30, 10-25, 10-20, 10-15, about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 consecutive modified sugars. In some embodiments, the first domain comprises two consecutive 2'-F modified sugars. In some embodiments, the first domain comprises three consecutive 2'-F modified sugars. In some embodiments, the first domain comprises four consecutive 2'-F modified sugars. In some embodiments, the first domain comprises five consecutive 2'-F modified sugars. In some embodiments, the first domain comprises six consecutive 2'-F modified sugars. In some embodiments, the first domain comprises 7 contiguous 2'-F modified sugars. In some embodiments, the first domain comprises 8 contiguous 2'-F modified sugars. In some embodiments, the first domain comprises 9 contiguous 2'-F modified sugars. In some embodiments, the first domain comprises 10 contiguous 2'-F modified sugars.In some embodiments, the first domain comprises two or more 2'-F modified sugar blocks, where each sugar in the 2'-F modified sugar block is independently a 2'-F modified sugar. In some embodiments, each 2'-F modified sugar block independently comprises or consists of 2, 3, 4, 5, 6, 7, 8, 9, or 10 contiguous 2'-F modified sugars as described herein. In some embodiments, two contiguous 2'-F modified sugar blocks are independently separated by a separation block, where the separation block comprises one or more sugars that are not independently 2'-F modified sugars. In some embodiments, each sugar in the separation block is independently not 2'-F modified. In some embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more) or all of the sugars in the separation block are independently not 2'-F modified. In some embodiments, the separation block comprises one or more bicyclic sugars (e.g., LNA sugars, cEt sugars, etc.), and / or one or more 2'-OR modified sugars (wherein R is an optionally substituted C. 1~6 In some embodiments, the separation block comprises one or more 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the two or more 2'-F unmodified sugars are contiguous. In some embodiments, the two or more 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the separation block is comprised of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more) 2'-OR modified sugars (wherein R is an optionally substituted C 1~6 In some embodiments, the separation block comprises two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more) contiguous 2'-ORF modified sugars (wherein R is an optionally substituted C1~6 In some embodiments, each 2'-OR modified sugar is independently a 2'-OMe sugar or a 2'-MOE sugar. In some embodiments, each 2'-OR modified sugar is independently a 2'-OMe sugar. In some embodiments, each 2'-OR modified sugar is independently a 2'-MOE sugar. In some embodiments, the separation block comprises one or more 2'-F modified sugars. In some embodiments, the 2'-F modified sugars in the separation block are not adjacent to one another. In some embodiments, the separation block does not comprise a 2'-F modified sugar. In some embodiments, each sugar in the separation block is independently a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, each sugar in each separation block is independently a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, each sugar in the separation block is independently a 2'-OR modified sugar (wherein R is an optionally substituted C 1~6 In some embodiments, each sugar in each separation block is independently a 2'-OR modified sugar, where R is an optionally substituted C 1~6 aliphatic). In some embodiments, each sugar in a separation block is independently a 2'-OMe or 2'-MOE modified sugar. In some embodiments, each sugar in each separation block is independently a 2'-OMe or 2'-MOE modified sugar. In some embodiments, each sugar in a separation block is independently a 2'-OMe modified sugar. In some embodiments, each sugar in a separation block is independently a 2'-MOE modified sugar. In some embodiments, a separation block comprises a 2'-OMe sugar and a 2'-MOE modified sugar. In some embodiments, each 2'-F block and each separation block independently comprises 1, 2, 3, 4, or 5 nucleosides. In some embodiments, each 2'-F block and each separation block independently comprises 1, 2, or 3 nucleosides.

[0182] In some embodiments, between about 5% and 100% (e.g., between about 10% and 100%, 20% and 100%, 30% and 100%, 40% and 100%, 50% and 80%, 50% and 85%, 50% and 90%, 50% and 95%, 60% and 80%, 60% and 85%, 60% and 90%, 60% and 95%, 60% and 100%, 65% and 80%, 65% and 85%, 65% and 90%, 65% and 95%, 65% and 100%, 70% and 80%, 70% and 85%, 70% and 90%, %, 70%-95%, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, etc., is independently a modified sugar. In some embodiments, between about 5% and 100% (e.g., between about 10% and 100%, 20% and 100%, 30% and 100%, 40% and 100%, 50% and 80%, 50% and 85%, 50% and 90%, 50% and 95%, 60% and 80%, 60% and 85%, 60% and 90%, 60% and 95%, 60% and 100%, 65% and 80%, 65% and 85%, 65% and 90%, 65% and 95%, 65% and 100%, 70% and 80%, 70% and 85%, 70% and 90%, 70%-95%, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% are independently 2'-F modified sugars. In some embodiments, the percentage is at least about 40%. In some embodiments, the percentage is at least about 50%. In some embodiments, the percentage is at least about 55%. In some embodiments, the percentage is at least about 60%.In some embodiments, the percentage is at least about 65%. In some embodiments, the percentage is at least about 70%. In some embodiments, the percentage is at least about 75%. In some embodiments, the percentage is at least about 80%. In some embodiments, the percentage is at least about 85%. In some embodiments, the percentage is at least about 90%. In some embodiments, the percentage is at least about 95%. In some embodiments, the percentage is about 100%. In some embodiments, the percentage is about 60% or about 5% or less. In some embodiments, the percentage is about 70% or about 70% or less. In some embodiments, the percentage is about 80% or about 80% or less. In some embodiments, the percentage is about 90% or about 90% or less.

[0183] In some embodiments, the first domain does not include a bicyclic sugar or a 2'-OR modified sugar where R is not -H. In some embodiments, the first domain includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) bicyclic sugars and / or 2'-OR modified sugars where R is not -H. In some embodiments, the first domain includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) 2'-OR modified sugars where R is not -H. In some embodiments, the first domain includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) 2'-OR modified sugars where R is optionally substituted C 1~10In some embodiments, the level of bicyclic sugars and / or 2'-OR modified sugars (where R is not -H), individually or combined, is relatively low compared to the level of 2'-F modified sugars. In some embodiments, the level of bicyclic sugars and / or 2'-OR modified sugars (where R is not -H), individually or combined, is about 10%-80% (e.g., about 10%-75%, 10%-70%, 10%-65%, 10%-60%, 10%-50%, about 20%-60%, about 30%-60%, about 20%-50%, about 30%-50%, about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60%, etc.). In some embodiments, the level of 2'-OR modified sugars (where R is not -H) (e.g., 2'-OMe and 2'-MOE modified sugars, if present, combined) is about 10%-70% (e.g., about 10%-60%, 10-50%, about 20-60%, about 30-60%, about 20-50%, about 30-50%, about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60%, etc.). In some embodiments, about 1% to 95% or less (e.g., about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, etc.) of the sugars in the first domain comprise 2'-OMe. In some embodiments, about 50% or less of the sugars in the first domain comprise 2'-OMe. In some embodiments, about 1% to 95% or less (e.g., about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, etc.) of the sugars in the first domain are 2′-OR, where R is an optionally substituted C 1~6 In some embodiments, no more than about 50% of the sugars in the first domain are 2'-OR, where R is an optionally substituted C 1~6 In some embodiments, no more than about 40% of the sugars in the first domain are 2'-OR, where R is an optionally substituted C 1~6In some embodiments, no more than about 30% of the sugars in the first domain are 2'-OR, where R is an optionally substituted C 1~6 In some embodiments, no more than about 25% of the sugars in the first domain are 2'-OR, where R is an optionally substituted C 1~6 In some embodiments, no more than about 20% of the sugars in the first domain are 2'-OR, where R is an optionally substituted C 1~6 In some embodiments, about 10% or less of the sugars in the first domain are 2'-OR, where R is an optionally substituted C 1~6 In some embodiments, the 2'-OR is 2'-MOE, as described herein. In some embodiments, the 2'-OR is 2'-MOE or 2'-OMe, as described herein. In some embodiments, the first domain comprises a 2'-N(R) 2 In some embodiments, the first domain comprises one or more (e.g., about 1-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.) modified sugars that include a modification. 2The first domain comprises one or more (e.g., about 1-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.) modified sugars comprising a modification. In some embodiments, the first domain comprises one or more (e.g., about 1-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.) bicyclic sugars (e.g., LNA sugars). In some embodiments, the first domain comprises one or more (e.g., about 1-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.) acyclic sugars (e.g., UNA sugars). In some embodiments, some of the 5'-terminal sugars in the first domain are independently 2'-OR modified sugars (where R is not -H). In some embodiments, some of the 5'-terminal sugars in the first domain (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) are independently 2'-OR modified sugars (where R is independently an optionally substituted C 1~6 In some embodiments, about the first 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) sugars from the 5' end of the first domain are independently 2'-OR modified sugars (wherein R is independently an optionally substituted C 1~6 In some embodiments, the first one is 2'-OR modified. In some embodiments, the first two are independently 2'-OR modified. In some embodiments, the first three are independently 2'-OR modified. In some embodiments, the first four are independently 2'-OR modified. In some embodiments, the first five are independently 2'-OR modified. In some embodiments, all 2'-OR modifications in a domain (e.g., a first domain), subdomain (e.g., a first subdomain), or oligonucleotide are the same. In some embodiments, the 2'-OR is 2'-MOE. In some embodiments, the 2'-OR is 2'-OMe.

[0184] In some embodiments, no sugar in the first domain comprises a 2'-OR. In some embodiments, no sugar in the first domain comprises a 2'-OMe. In some embodiments, no sugar in the first domain comprises a 2'-MOE. In some embodiments, no sugar in the first domain comprises a 2'-MOE or a 2'-OMe. In some embodiments, no sugar in the first domain comprises a 2'-OR (wherein R is optionally substituted C 1~6 In some embodiments, each sugar in the first domain does not include a 2'-F.

[0185] In some embodiments, about 40-70% (e.g., about 40%-70%, 40%-60%, 50%-70%, 50%-60%, etc., or about 40%, 45%, 50%, 55%, 60%, 65%, 70%, etc.) of the sugars in the first domain are 2'-F modified and about 10%-60% (e.g., about 10%-50%, 20%-60%, 30%-60%, 30%-50%, 40%-50%, etc., or about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60%) of the sugars in the first domain are, independently, 2'-OR modified (where R is not -H) or are bicyclic sugars (e.g., LNA sugars, cEt sugars, etc.). In some embodiments, between about 20% and 60% of the sugars in the first domain are 2'-F modified. In some embodiments, between about 25% and 60% of the sugars in the first domain are 2'-F modified. In some embodiments, between about 30% and 60% of the sugars in the first domain are 2'-F modified. In some embodiments, between about 35% and 60% of the sugars in the first domain are 2'-F modified. In some embodiments, between about 40% and 60% of the sugars in the first domain are 2'-F modified. In some embodiments, between about 50% and 60% of the sugars in the first domain are 2'-F modified. In some embodiments, between about 50% and 70% of the sugars in the first domain are 2'-F modified. In some embodiments, between about 20% and 60% of the sugars in the first domain are independently 2'-OR modified (wherein R is not -H) or are bicyclic sugars. In some embodiments, about 30%-60% of the sugars in the first domain are independently 2'-OR modified (where R is not -H) or are bicyclic sugars. In some embodiments, about 40%-60% of the sugars in the first domain are independently 2'-OR modified (where R is not -H) or are bicyclic sugars. In some embodiments, about 30%-50% of the sugars in the first domain are independently 2'-OR modified (where R is not -H) or are bicyclic sugars. In some embodiments, about 40%-50% of the sugars in the first domain are independently 2'-OR modified (where R is not -H) or are bicyclic sugars.In some embodiments, each of the sugars in the first domain that are independently 2'-R modified (where R is not -H) or are bicyclic sugars is independently a 2'-OR modified sugar (where R is not -H). In some embodiments, each of these is independently a 2'-OR modified sugar (where R is C. 1~6 In some embodiments, each of these is independently a 2'-OR modified sugar (formula, R is C 1~6 In some embodiments, each of these is independently a 2'-OMe or a 2'-MOE modified sugar.

[0186] In some embodiments, a first domain is 5' to a second domain, and one or more sugars (e.g., 1-5, 1-3, 1, 2, 3, 4, 5, etc.) first from the 5' end of the first domain are each independently a 2'-OR modified sugar or a bicyclic sugar, and R is not -H (e.g., optionally substituted C 1~6 In some embodiments, the first domain is 5' to the second domain, and the first three sugars from the 5' end of the first domain are each independently a 2'-OR modified sugar or a bicyclic sugar, and R is not -H (e.g., optionally substituted C 1~6 alkyl). In some embodiments, each such sugar is independently a 2'-OMe or 2'-MOE modified sugar. In some embodiments, each such sugar is independently a 2'-OMe modified sugar. In some embodiments, each such sugar is independently a 2'-MOE modified sugar.

[0187] In some embodiments, the first domain comprises about 1 to 50 (e.g., about 5, 6, 7, 8, 9, or 10 to about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, etc., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, etc.) modified internucleotide linkages. In some embodiments, between about 5% and 100% (e.g., between about 10% and 100%, 20% and 100%, 30% and 100%, 40% and 100%, 50% and 80%, 50% and 85%, 50% and 90%, 50% and 95%, 60% and 80%, 60% and 85%, 60% and 90%, 60% and 95%, 60% and 100%, 65% and 80%, 65% and 85%, 65% and 90%, 65% and 95%, 65% and 100%, 70% and 80%, 70% and 85%, 70% and 90 ... 0%, 70%-95%, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% (e.g., 0%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%) is a modified internucleotide linkage. In some embodiments, each internucleotide linkage in the first domain is independently a modified internucleotide linkage. In some embodiments, each modified internucleotide linkage is independently a chiral internucleotide linkage. In some embodiments, the modified or chiral internucleotide linkage is a phosphorothioate internucleotide linkage. In some embodiments, the modified or chiral internucleotide linkage is a non-negatively charged internucleotide linkage. In some embodiments, the modified or chiral internucleotide linkage is a neutral internucleotide linkage (e.g., n001).In some embodiments, each modified internucleotide linkage is independently a phosphorothioate internucleotide linkage or a non-negatively charged internucleotide linkage. In some embodiments, each modified internucleotide linkage is independently a phosphorothioate internucleotide linkage or a neutral internucleotide linkage. In some embodiments, each modified internucleotide linkage is independently a phosphorothioate internucleotide linkage. In some embodiments, at least about 1 to 50 (e.g., about 5, 6, 7, 8, 9, or 10 to about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, etc., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, etc.) chiral internucleotide linkages in the first domain are chiral controlled. In some embodiments, at least 5% to 100% (e.g., about 10% to 100%, 20 to 100%, 30% to 100%, 40% to 100%, 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65% to 100%, 70% to 80%, 70% to 80%, 8 ... 5%, 70%-90%, 70%-95%, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, etc., are chiral controlled.In some embodiments, at least 5% to 100% (e.g., about 10% to 100%, 20 to 100%, 30% to 100%, 40% to 100%, 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65% to 100%, 70% to 8 ... % to 85%, 70% to 90%, 70% to 95%, 70% to 100%, 75% to 80%, 75% to 85%, 75% to 90%, 75% to 95%, 75% to 100%, 80% to 85%, 80% to 90%, 80% to 95%, 80% to 100%, 85% to 90%, 85% to 95%, 85% to 100%, 90% to 95%, 90% to 100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% (e.g., each independently is chiral controlled). In some embodiments, at least about 1 to 50 (e.g., about 5, 6, 7, 8, 9, or 10 to about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, etc., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, etc.) chiral internucleotide linkages in the first domain are Sp.In some embodiments, at least about 1 to 50 (e.g., about 5, 6, 7, 8, 9, or 10 to about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, such as about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, such as about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) phosphorothioate internucleotide linkages in the first domain are Sp. In some embodiments, at least 5% to 100% (e.g., about 10% to 100%, 20 to 100%, 30% to 100%, 40% to 100%, 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65% to 100%, 70% to 80%, 70% to 85%, 70%-90%, 70%-95%, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, etc.) is Sp.In some embodiments, at least 5% to 100% (e.g., about 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65% to 100%, 70% to 8 ... 0%-85%, 70%-90%, 70%-95%, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% (e.g., 0%-85%, 70%-90%, 70%-95%, 80%-100%, 7 ... In some embodiments, this number is 4 or greater. In some embodiments, this number is 5 or greater. In some embodiments, this number is 6 or greater. In some embodiments, this number is 7 or greater. In some embodiments, this number is 8 or greater. In some embodiments, this number is 9 or greater. In some embodiments, this number is 10 or greater. In some embodiments, this number is 11 or greater. In some embodiments, this number is 12 or greater. In some embodiments, this number is 13 or greater. In some embodiments, this number is 14 or greater. In some embodiments, this number is 15 or greater. In some embodiments, the percentage is at least about 50%. In some embodiments, the percentage is at least about 55%. In some embodiments, the percentage is at least about 60%. In some embodiments, the percentage is at least about 65%. In some embodiments, the percentage is at least about 70%.In some embodiments, the percentage is at least about 75%. In some embodiments, the percentage is at least about 80%. In some embodiments, the percentage is at least about 85%. In some embodiments, the percentage is at least about 90%. In some embodiments, the percentage is at least about 95%. In some embodiments, the percentage is about 100%. In some embodiments, each internucleotide linkage linking two first domain nucleosides is independently a modified internucleotide linkage. In some embodiments, each modified internucleotide linkage is independently a chiral internucleotide linkage. In some embodiments, each modified internucleotide linkage is independently a phosphorothioate internucleotide linkage. In some embodiments, each chiral internucleotide linkage is independently a phosphorothioate internucleotide linkage. In some embodiments, each modified internucleotide linkage is independently a Sp chiral internucleotide linkage. In some embodiments, each modified internucleotide linkage is independently a phosphorothioate internucleotide linkage of Sp. In some embodiments, each chiral internucleotide linkage is independently a phosphorothioate internucleotide linkage of Sp. In some embodiments, the internucleotide linkage of the first domain is linked to two nucleosides of the first domain. In some embodiments, the internucleotide linkages linked to the nucleosides in the first domain and the nucleosides in the second domain can be properly considered to be the internucleotide linkage of the first domain. In some embodiments, the nucleosides in the first domain and the nucleosides in the second domain can be properly considered to be the internucleotide linkage of the first domain. The internucleotide linkage attached to the side is a modified internucleotide linkage; in some embodiments, it is a chiral internucleotide linkage; in some embodiments, it is chiral controlled; in some embodiments, it is Rp; in some embodiments, it is Sp. In many embodiments, a high percentage of Sp internucleotide linkages (e.g., compared to Rp internucleotide linkages and / or natural phosphate linkages) results in improved properties and / or activity (e.g., increased stability and / or increased adenosine editing activity).

[0188] In some embodiments, the first domain comprises a certain level of Rp internucleotide linkages. In some embodiments, the level may be about, e.g., about 5%-100%, about 10%-100%, 20%-100%, 30%-100%, 40%-100%, 50%-80%, 50%-85%, 50%-90%, 50%-95%, 60%-80%, 60%-85%, 60%-90%, 60%-95%, 60%-100%, 65%-80%, 65%-85%, 65%-90%, 65%-95%, 65%-100%, 70%-80%, 70%~85%, 70%~90%, 70%~95%, 70%~100%, 75%~80%, 75%~85%, 75%~90%, 75%~95%, 75%~100%, 80%~85%, 80%~90%, 80%~95%, 80%~100%, 85%~90%, 85%~95%, 85%~100%, 90%~95%, 90%~100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the level is about, e.g., about 5% to 100%, about 10% to 100%, 20 to 100%, 30% to 100%, 40% to 100%, 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65% to 100%, 70% to 80%, %, 70%~85%, 70%~90%, 70%~95%, 70%~100%, 75%~80%, 75%~85%, 75%~90%, 75%~95%, 75%~100%, 80%~85%, 80%~90%, 80%~95%, 80%~100%, 85%~90%, 85%~95%, 85%~100%, 90%~95%, 90%~100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.In some embodiments, the level is about, e.g., about 5% to 100%, about 10% to 100%, 20 to 100%, 30% to 100%, 40% to 100%, 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 10 ... 80%, 70%-85%, 70%-90%, 70%-95%, 70%-100%, 75%-80%, 75%-85%, 75%-90%, 75%-95%, 75%-100%, 80%-85%, 80%-90%, 80%-95%, 80%-100%, 85%-90%, 85%-95%, 85%-100%, 90%-95%, 90%-100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, etc. In some embodiments, the percentage is about 50% or less than about 50%. In some embodiments, the percentage is at l...

Claims

1. The first domain is a first domain whose length consists of approximately 10 or more nucleic acid bases; A second domain having a length of approximately 10 or more nucleic acid bases; and 2'-F modified sugar; Includes, The base sequence of the first domain is complementary to the first portion of the base sequence of the target nucleic acid; The base sequence of the second domain is complementary to the second portion of the base sequence of the target RNA nucleic acid; The first and second portions of the base sequence of the target RNA nucleic acid are separated in the target nucleic acid by gaps of about or at least about 50 nucleic acid bases. The length of an oligonucleotide is approximately 25 to 50 nucleic acid bases. Oligonucleotides.

2. The first domain; and Second domain Includes, The first domain is characterized by being able to form a double helix with the first portion of the target nucleic acid; The second domain is characterized by being able to form a double helix with a second portion of the target nucleic acid; An oligonucleotide in which the first and second portions of the base sequence of the target nucleic acid are separated by a gap.

3. The oligonucleotide is 5'-N １ N ０ N －１ -3' (in the formula, N －１ , N ０ , and N １ Each of them is independently a nucleoside, and N ０ The nucleic acid base is BA, and BA is 【Chemistry 1】 (is) Includes, Optionally, the oligonucleotide comprises 5'-N １ N ０ N －１ -3', where each of N －１ N ０ and N １ is independently a nucleoside; the nucleobase of N ０ is BA, where BA comprises ring BA or a tautomer thereof, and ring BA has the formula BA-III-e: 【Chemistry 2】 BA-III-e (In the formula, X １ is -N(-)- or -C(-)=; W Ｘ２ and W Ｘ６ These are, independently, O, S, or Se; R Ｂ４ These are halogen, -CN, -NO ２ , or -L Ｂ４ -R Ｂ４１ (In the formula, R Ｂ４１ (is R'); R Ｂ５ These are halogen, -CN, -NO ２ , or -L Ｂ５ -R Ｂ５１ (In the formula, R Ｂ５１ is -R', -N(R') ２ (which is -OR', or -SR') L Ｂ４ and L Ｂ５ Each of them is independent of L Ｂ And; Each L Ｂ These are independently, covalently bonded, or optionally substituted divalent carbon atoms having 0 to 6 heteroatoms. １～１０ A saturated or partially unsaturated chain, in which one or more methylene units are optionally and independently -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O) ２ -, -S(O) ２ Replaced with N(R')-, -C(O)S-, or -C(O)O-; Each -Cy- is independently an optionally substituted 3-20 member monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms; Each R' is independently -R, -C(O)R, -C(O)OR, -C(O)N(R) ２ , or -SO ２ It is R; Each R is independently -H or C １～２０ Aliphatic carbon atoms containing 1 to 10 heteroatoms １～２０ Heteroliphatic, C ６～２０ Ariel, C ６～２０ Aryl aliphatic carbon atoms containing 1 to 10 heteroatoms ６～２０ The group is optionally substituted from aryl heteroaliphatic groups, 5-20 membered heteroaryl groups having 1-10 heteroatoms, and 3-20 membered heterocyclines having 1-10 heteroatoms, or: Two R groups are combined either arbitrarily and independently to form a covalent bond, or: Two or more R groups on the same atom, optionally and independently, together with the atom, form an optionally substituted 3-20 member monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms in addition to the atom; or The structure has two or more R groups on two or more atoms, which are arbitrarily and independently combined with the intervening atoms to form an arbitrarily substituted 3-30 member monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms in addition to the intervening atoms. Optionally, N ０ The aforementioned sugar is 2'-OR modified (wherein R is optionally substituted C). １～６ (Includes aliphatic) Selectively, the oligonucleotide is 5'-N １ N ０ N －１ -3' (in the formula, N －１ , N ０ , and N １ Each of them is independently a nucleoside, and N ０ The nucleic acid base is BA, and BA is 【Transformation 3】 (is) including, Selectively, the oligonucleotide is 5'-N １ N ０ N －１ -3' is included, N －１ , N ０ , and N １ Each of them is an independent nucleoside; N ０ The nucleic acid base is BA, and BA includes ring BA or its tautomer, and ring BA is of formula BA-VI: 【Chemistry 4】 BA-VI (In the formula, each 【Transformation 5】 These are, independently, single or double bonds; X １’ is -N(-)- or -C(-)=; X ２’ is -C(W Ｘ２’ )-,-C(R Ｂ２’ ) = -C(OR Ｂ２’ )=, -N=, or optionally substituted -CH= or -CH ２ - (wherein, R Ｂ２’ These are halogen, -CN, -NO ２ , or -L Ｂ２’ -R' and W Ｘ２’ (is O, S, or Se); X ３’ is -N(R Ｂ３’ )=, -N=, -C(R Ｂ３’ ) = or optionally substituted -NH- or -CH=, R Ｂ３’ These are halogen, -CN, -NO ２ , or -L Ｂ３’ -R'; X ４’ is -C(R Ｂ４’ )=, -C(OR Ｂ４’ )=, -C(-N(R Ｂ４’ )) ２ =, -C(R Ｂ４’ ) ２ -, -C(W Ｘ４’ )-, -C(=NR Ｂ４’ )-, -N(R Ｂ４’ )-, -N=, or optionally substituted -CH=, -NH- or -CH ２ -(where each R Ｂ４’ is independently halogen, -CN, -NO ２ , or -L Ｂ４’ -R Ｂ４１’ or two Rs on the same atom are combined to form =O, =C(-L Ｂ４’ -R Ｂ４’ ) Ｂ４１’ ), =N-L ２ -R Ｂ４’ or optionally substituted =CH Ｂ４１’ or =NH (where each R ２ is independently -R'), and W Ｂ４１’ is O, S or Se);​​ X ５’ is -C(R Ｂ５’ ) ２ -, -N(R Ｂ５’ )-,-C(R Ｂ５’ ) = -C(W Ｘ５’ )-, -N=, or optionally substituted -NH-, -CH ２ - or -CH = (where each R Ｂ５’ These are, independently, halogen, -CN, and -NO ２ , or -L Ｂ５’ -R Ｂ５１’ (In the formula, R Ｂ５１’ is -R', -N(R') ２ (which is -OR', or -SR') and W Ｘ５’ (is O, S, or Se); X ６’ is -C(R Ｂ６’ ) = -C(OR Ｂ６’ ) = -C(R Ｂ６’ ) ２ -, -C(W Ｘ６’ )-,-C(-N(R Ｂ６’ ) ２ ) =, -N = or optionally substituted -NH-, -CH ２ - or -CH = (where each R Ｂ６’ These are, independently, halogen, -CN, and -NO ２ , or -L Ｂ６’ -R Ｂ６１’ or two R on the same atom Ｂ６’ Combining them, =O, =C(-L) Ｂ６’ -R Ｂ６１’ ) ２ , = N - L Ｂ６’ -R Ｂ６１’ , or optionally substituted =CH ２ Or = NH (where R Ｂ６１’ It independently forms R', and W Ｘ６’ (is O, S, or Se); X ７’ is -C(R Ｂ７’ ) = -C(OR Ｂ７’ ) = -C(R Ｂ７’ ) ２ -, -C(W Ｘ７’ )-,-C(-N(R Ｂ７’ ) ２ ) = -N(R Ｂ７’ )-, -N=, or optionally substituted -NH-, -CH ２ - or -CH = (where each R Ｂ７’ These are, independently, halogen, -CN, and -NO ２ , or -L Ｂ７’ -R Ｂ７１’ or two R on the same atom Ｂ７’ Combining them, =O, =C(-L) Ｂ７’ -R Ｂ７１’ ) ２ , = N - L Ｂ７’ -R Ｂ７１’ , or optionally substituted =CH ２ Or = NH (where R Ｂ７１’ It independently forms R', and W Ｘ７’ (is O, S, or Se); X ８’ and X ９’ Each of them is independently either C or N; L Ｂ２’ , L Ｂ３’ , L Ｂ４’ , L Ｂ５’ , L Ｂ６’ and L Ｂ７’ Each of them is independent of L Ｂ And; Each L Ｂ These are independently, covalently bonded, or optionally substituted divalent carbon atoms having 0 to 6 heteroatoms. １～１０ A saturated or partially unsaturated chain, in which one or more methylene units are optionally and independently -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O) ２ -, -S(O) ２ Replaced with N(R')-, -C(O)S-, or -C(O)O-; Each -Cy- is independently an optionally substituted 3-20 member monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms; Each R' is independently -R, -C(O)R, -C(O)OR, -C(O)N(R) ２ , or -SO ２ It is R; Each R is independently -H or C １～２０ Aliphatic carbon atoms containing 1 to 10 heteroatoms １～２０ Heteroliphatic, C ６～２０ Ariel, C ６～２０ Aryl aliphatic carbon atoms containing 1 to 10 heteroatoms ６～２０ The group is optionally substituted from aryl heteroaliphatic groups, 5-20 membered heteroaryl groups having 1-10 heteroatoms, and 3-20 membered heterocyclines having 1-10 heteroatoms, or: Two R groups are combined either arbitrarily and independently to form a covalent bond, or: Two or more R groups on the same atom, optionally and independently, together with the atom, form an optionally substituted 3-20 member monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms in addition to the atom; or The oligonucleotide according to claim 1 or 2, having a structure in which two or more R groups on two or more atoms are optionally and independently combined with the intervening atoms to form an optionally substituted 3-30 member monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms in addition to the intervening atoms.

4. The length of the first domain is approximately 10 to 30 nucleic acid bases. The length of the second domain is optionally set to approximately 10 to 30 nucleic acid bases. The oligonucleotide according to any one of claims 1 to 3, wherein the length of the gap is optionally about or at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, 60, 70, 80, 90, or 100 nucleic acid bases.

5. The first domain is connected to the second domain via a linker, and optionally the linker is or contains an oligonucleotide ("linker oligonucleotide"), Selectively, the complementarity of the linker oligonucleotide to the gap in the target nucleic acid is approximately 30%, 25%, 20%, 10%, or 5% or less. The oligonucleotide according to any one of claims 1 to 4, wherein the linker optionally comprises polyvinyl ether, polyethylene, polypropylene, polyethylene glycol (PEG), polypropylene glycol (PEG), polyvinyl alcohol (PVA), polyglycolide (PGA), polylactide (PLA), polycaprolactone (PCL), or copolymers thereof.

6. The aforementioned target nucleic acid is mRNA. When the oligonucleotide comes into contact with a target nucleic acid containing target adenosine in the system, the target adenosine in the target nucleic acid is modified. Selectively, the target adenosine is a mutation from guanine. Selectively, the number of oligonucleotides is approximately 10 to 200 (for example, approximately 10 to 20, 10 to 30, 10 to 40, 10 to 50, 10 to 60, 10 to 70, 10 to 80, 10 to 90, 10 to 100, 10 to 120, 10 to 150, 20 to 30, 20 to 40, 20 to 50, 20 to 60, 20 to 70, 20 to 80, 20 to 90, 20 to 100, 20 to 120, 20 to 150, 20 to 200, 25 to 30, 25 to 40, 25 to 50, 2 Nucleic acid bases have lengths of 5-60, 25-70, 25-80, 25-90, 25-100, 25-120, 25-150, 25-200, 30-40, 30-50, 30-60, 30-70, 30-80, 30-90, 30-100, 30-120, 30-150, 30-200, 10, 20, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 45, 50, 60, etc. Selectively, the oligonucleotide has a length of approximately 30 to 40 nucleic acid bases, The first domain has a length of approximately 2 to 50 nucleic acid bases (for example, approximately 5, 6, 7, 8, 9, or 10 to approximately 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50 bases, or approximately 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50 bases, etc.), Selectively, at least about 1 to 50 (for example, about 5, 6, 7, 8, 9, or 10 to about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50, etc., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, etc.) chiral nucleotide interlinks in the first subdomain are chiral controlled. The second domain has a length of approximately 2 to 50 nucleic acid bases (for example, approximately 5, 6, 7, 8, 9, or 10 to approximately 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50 bases, or approximately 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 40, or 50 bases, etc.), If the oligonucleotide is optionally aligned with the target nucleic acid in terms of complementarity, the second domain includes one or more fluctuation pairs (e.g., 1 to 10, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.), If the oligonucleotide is optionally aligned with the target nucleic acid in terms of complementarity, the second domain comprises the nucleoside opposite to the target adenosine. The oligonucleotide according to any one of claims 1 to 5, wherein, if the oligonucleotide is optionally aligned with a target nucleic acid in terms of complementarity, the first and second domains each independently comprise a nucleoside opposite to the target adenosine.

7. The first and second domains each independently contain a nucleic acid base opposite to the target adenosine, each opposite nucleic acid base containing the nucleic acid base BA, where BA is a ring BA or a tautomer thereof, or contains the ring BA having the formula BA-III-e: 【Transformation 6】 BA-III-e (In the formula, X １ is -N(-)- or -C(-)=; W Ｘ２ and W Ｘ６ Each of these is independently O, S, or Se; R Ｂ４ These are halogen, -CN, -NO ２ , or -L Ｂ４ -R Ｂ４１ (In the formula, R Ｂ４１ (is R'); R Ｂ５ These are halogen, -CN, -NO ２ , or -L Ｂ５ -R Ｂ５１ (In the formula, R Ｂ５１ is -R', -N(R') ２ (which is -OR', or -SR') L Ｂ４ and L Ｂ５ Each of them is independently L; Each L Ｂ These are independently, covalently bonded, or optionally substituted divalent carbon atoms having 0 to 6 heteroatoms. １～１０ A saturated or partially unsaturated chain, in which one or more methylene units are optionally and independently -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O) ２ -, -S(O) ２ Replaced with N(R')-, -C(O)S-, or -C(O)O-; Each -Cy- is independently an optionally substituted 3-20 member monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms; Each R' is independently -R, -C(O)R, -C(O)OR, -C(O)N(R) ２ , or -SO ２ It is R; Each R is independently -H or C １～２０ Aliphatic carbon atoms containing 1 to 10 heteroatoms １～２０ Heteroliphatic, C ６～２０ Ariel, C ６～２０ Aryl aliphatic carbon atoms containing 1 to 10 heteroatoms ６～２０ The group is optionally substituted from aryl heteroaliphatic groups, 5-20 membered heteroaryl groups having 1-10 heteroatoms, and 3-20 membered heterocyclines having 1-10 heteroatoms, or: Two R groups are combined either arbitrarily and independently to form a covalent bond, or: Two or more R groups on the same atom, optionally and independently, together with the atom, form an optionally substituted 3-20 member monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms in addition to the atom; or The oligonucleotide according to any one of claims 1 to 6, having a structure in which two or more R groups on two or more atoms are optionally and independently combined with the intervening atoms to form an optionally substituted 3 to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.

8. The first and second domains each independently contain a nucleic acid base opposite to the target adenosine, each opposite nucleic acid base containing the nucleic acid base BA, where BA is or contains a ring BA or a tautomer thereof, and the ring BA has the formula BA-VI: 【Transformation 7】 BA-VI (In the formula, each 【Transformation 8】 These are, independently, single or double bonds; X １’ is -N(-)- or -C(-)=; X ２’ is -C(W Ｘ２’ )-,-C(R Ｂ２’ ) = -C(OR Ｂ２’ )=, -N=, or optionally substituted -CH= or -CH ２ - (wherein, R Ｂ２’ These are halogen, -CN, -NO ２ , or -L Ｂ２’ -R' and W Ｘ２’ (is O, S, or Se); X ３’ is -N(R Ｂ３’ )=, -N=, -C(R Ｂ３’ ) = or optionally substituted -NH- or -CH=, R Ｂ３’ These are halogen, -CN, -NO ２ , or -L Ｂ３’ -R'; X ４’ is -C(R Ｂ４’ ) = -C(OR Ｂ４’ )=, -C(-N(R Ｂ４’ ) ２ ) = -C(R Ｂ４’ ) ２ -, -C(W Ｘ４’ )-,-C(=NR Ｂ４’ )-,-N(R Ｂ４’ )-, -N=, or optionally substituted -CH=, -NH- or -CH ２ -(In the formula, each R Ｂ４’ These are, independently, halogen, -CN, and -NO ２ , or -L Ｂ４’ -R Ｂ４１’ or two R on the same atom Ｂ４’ Combining them, =O, =C(-L) Ｂ４’ -R Ｂ４１’ ) ２ , = N - L Ｂ４’ -R Ｂ４１’ , or optionally substituted =CH ２ Or = NH (where R Ｂ４１’ These independently form (which is -R') and W Ｘ４’ (is O, S, or Se); X ５’ is -C(R Ｂ５’ ) ２ -, -N(R Ｂ５’ )-,-C(R Ｂ５’ ) = -C(W Ｘ５’ )-, -N=, or optionally substituted -NH-, -CH ２ - or -CH = (where each R Ｂ５’ These are, independently, halogen, -CN, and -NO ２ , or -L Ｂ５’ -R Ｂ５１’ (In the formula, R Ｂ５１’ is -R', -N(R') ２ (which is -OR', or -SR') and W Ｘ５’ (is O, S, or Se); X ６’ is -C(R Ｂ６’ ) = -C(OR Ｂ６’ ) = -C(R Ｂ６’ ) ２ -, -C(W Ｘ６’ )-,-C(-N(R Ｂ６’ ) ２ ) =, -N = or optionally substituted -NH-, -CH ２ - or -CH = (where each R Ｂ６’ These are, independently, halogen, -CN, and -NO ２ , or -L Ｂ６’ -R Ｂ６１’ or two R on the same atom Ｂ６’ Combining them, =O, =C(-L) Ｂ６’ -R Ｂ６１’ ) ２ , = N - L Ｂ６’ -R Ｂ６１’ , or optionally substituted =CH ２ Or = NH (where R Ｂ６１’ It independently forms R', and W Ｘ６’ (is O, S, or Se); X ７’ is -C(R Ｂ７’ ) = -C(OR Ｂ７’ ) = -C(R Ｂ７’ ) ２ -, -C(W Ｘ７’ )-,-C(-N(R Ｂ７’ ) ２ ) = -N(R Ｂ７’ )-, -N=, or optionally substituted -NH-, -CH ２ - or -CH = (where each R Ｂ７’ These are, independently, halogen, -CN, and -NO ２ , or -L Ｂ７’ -R Ｂ７１’ or two R on the same atom Ｂ７’ Combining them, =O, =C(-L) Ｂ７’ -R Ｂ７１’ ) ２ , = N - L Ｂ７’ -R Ｂ７１’ , or optionally substituted =CH ２ Or = NH (where R Ｂ７１’ It independently forms R', and W Ｘ７’ (is O, S, or Se); X ８’ and X ９’ Each of them is independently either C or N; L Ｂ２’ , L Ｂ３’ , L Ｂ４’ , L Ｂ５’ , L Ｂ６’ and L Ｂ７’ Each of them is independent of L Ｂ And; Each L Ｂ These are independently, covalently bonded, or optionally substituted divalent carbon atoms having 0 to 6 heteroatoms. １～１０ A saturated or partially unsaturated chain, in which one or more methylene units are optionally and independently -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O) ２ -, -S(O) ２ Replaced with N(R')-, -C(O)S-, or -C(O)O-; Each -Cy- is independently an optionally substituted 3-20 member monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms; Each R' is independently -R, -C(O)R, -C(O)OR, -C(O)N(R) ２ , or -SO ２ It is R; Each R is independently -H or C １～２０ Aliphatic carbon atoms containing 1 to 10 heteroatoms １～２０ Heteroliphatic, C ６～２０ Ariel, C ６～２０ Aryl aliphatic carbon atoms containing 1 to 10 heteroatoms ６～２０ The group is optionally substituted from aryl heteroaliphatic groups, 5-20 membered heteroaryl groups having 1-10 heteroatoms, and 3-20 membered heterocyclines having 1-10 heteroatoms, or: Two R groups are combined either arbitrarily and independently to form a covalent bond, or: Two or more R groups on the same atom, optionally and independently, together with the atom, form an optionally substituted 3-20 member monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms in addition to the atom; or The oligonucleotide according to any one of claims 1 to 7, having a structure in which two or more R groups on two or more atoms are optionally and independently combined with the intervening atoms to form an optionally substituted 3 to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.

9. The sugar of the nucleoside containing the nucleic acid base on the opposite side is 2'-OR modified (wherein R is optionally substituted C). １～６ (Includes aliphatic) The oligonucleotide according to any one of claims 1 to 8, wherein the sugar of the nucleoside containing the opposite nucleic acid base optionally includes a natural DNA sugar.

10. A method for preparing an oligonucleotide or composition, comprising coupling the -OH group of an oligonucleotide or nucleoside with a phosphoramidite, wherein the nucleic acid base of the phosphoramidite is any one of the nucleic acid bases of claims 1 to 9 or a tautomer thereof, and the nucleic acid base or tautomer thereof is optionally substituted or protected. Optionally, the phosphoramidite is 【Chemistry 9】 Or a method having the structure of a salt thereof.

11. A method for deamination of target adenosine in a target nucleic acid, comprising contacting the target nucleic acid with an oligonucleotide or composition according to any one of claims 1 to 10, wherein the oligonucleotide targets the target adenosine.

12. A method for producing, restoring, or increasing the level of a specific nucleic acid or its product, comprising contacting a target nucleic acid with an oligonucleotide or composition according to any one of claims 1 to 11, wherein the target nucleic acid comprises a target adenosine, and the specific nucleic acid differs from the target nucleic acid in that it has I or G instead of the target adenosine.

13. A method comprising contacting an oligonucleotide or composition according to any one of claims 1 to 12 with a sample containing a target nucleic acid and adenosine deaminase, The base sequence of the oligonucleotide in the oligonucleotide composition is substantially complementary to that of the target nucleic acid; The target nucleic acid contains the target adenosine; A method for modifying target adenosine.

14. The method comprises obtaining a first level of modification of target adenosine in the target nucleic acid (the level being observed when the first oligonucleotide composition comes into contact with a sample containing the target nucleic acid and adenosine deaminase, and the first oligonucleotide composition comprises a plurality of first oligonucleotides sharing the same nucleotide sequence substantially complementary to that of the target nucleic acid); The first level of modification of the target adenosine is higher than the reference level of modification of the target adenosine, the reference level being observed when the reference oligonucleotide composition is in contact with a sample containing the target nucleic acid and adenosine deaminase, and the reference oligonucleotide composition comprises a plurality of reference oligonucleotides that share the same base sequence substantially complementary to that of the target nucleic acid; The first plurality of oligonucleotides comprises one or more chirally controlled chiral nucleotide bonds; A method wherein the reference oligonucleotides described above do not contain chiral-controlled chiral nucleotide bonds (the reference oligonucleotide composition is a "sterically disordered" composition).

15. An oligonucleotide or composition according to any one of claims 1 to 14 for preventing or treating a condition, disorder, or disease associated with a G-to-A mutation, wherein the oligonucleotide targets a G-to-A mutation for editing, and the prevention or treatment comprises administering or delivering an effective amount of the oligonucleotide or composition to a subject susceptible to the condition.

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