Oligonucleotide compositions and uses thereof

Abstract

Among other things, the present disclosure provides oligonucleotides comprising various patterns of modifications, e.g., sugar modifications, internucleotidic linkage modifications. Also provided herein are compositions comprising such oligonucleotides. Methods of making and using oligonucleotides comprising various patterns of modifications are further provided.

Description

Attorney Docket No.: 2010794-3211OLIGONUCLEOTIDE COMPOSITIONS AND USES THEREOFCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to United States Provisional Application Nos. 63 / 773,358, filed March 17, 2025, 63 / 773,403, filed March 17, 2025, 63 / 906,205, filed October 27, 2025, and 63 / 906,207, filed October 27, 2025, the entirety of each of which is incorporated herein by reference.BACKGROUND

[0002] Modulation of expression of certain genes, transcripts, or products encoded thereby (e.g., polypeptides) can occur through usage of modified oligonucleotides. Oligonucleotide modifications including various sugar modifications, nucleobase modifications, and internucleotidic linkage modifications can be used to alter properties (e.g., potency, stability) of oligonucleotides. Due to variation in oligonucleotide modifications and structures and properties thereof, various patterns of modifications may be implemented to provide desired properties (e.g., increased activities) to oligonucleotides.SUMMARY

[0003] Oligonucleotides can be used to modulate gene expression to provide treatments for various conditions, disorders, and diseases. Chemical modifications of oligonucleotides have been developed to provide, e.g., increased potency and / or increased stability. Such chemical modifications include sugar modifications (e.g., 2’-modified sugars (e.g., 2’-F, 2’-0Me, 2’-M0E), GNA sugars, LNA sugars, UNA sugars), nucleobases modifications (e.g., pseudouridine, 5-methylcytidine), and internucleotidic linkage modifications (e.g., phosphorothioate internucleotidic linkages, methylphosphonate internucleotidic linkages, chirally controlled internucleotidic linkages). In view of the various possible chemical modifications, a large variety of chemical modifications can be implemented in combination in any particular oligonucleotide.

[0004] Various patterns of oligonucleotide modifications (e.g., sugar modifications, nucleobase modifications, and / or internucleotidic linkage modifications) can be utilized to provide desired properties and / or activities of an oligonucleotide. A pattern of modifications can be used across various base sequences and target transcripts. Certain patterns of modifications may be utilized in certain types of oligonucleotides, e.g., RNAi oligonucleotides, siRNA oligonucleotides.

[0005] Among other things, the present disclosure provides technologies (e.g., oligonucleotides, compositions, methods, etc.) for modulating expression of various target transcripts. The present disclosure provides the recognition that certain patterns of modifications (e.g., sugar modifications, internucleotidic linkage modifications) are particularly useful in inhibiting expression of various target transcripts.Attorney Docket No.: 2010794-3211Modification patterns provided herein can be utilized with oligonucleotides targeting any number of different target transcripts and with oligonucleotides of various modalities, e.g., RNAi, e.g., siRNA. Oligonucleotides provided herein can be utilized, for example, in methods of treating a disease or disorder associated with aberrant expression of various target transcripts.

[0006] In some embodiments, the present disclosure provides oligonucleotides, or a pharmaceutically acceptable salt thereof, comprising a sense strand, an antisense strand, or any combination thereof. In some embodiments, a sense strand and / or an antisense strand comprises a pattern of modifications, e.g., sugar modifications. In some embodiments, an oligonucleotide is a pharmaceutically acceptable salt.

[0007] In some embodiments, an antisense strand comprises mN fN (mN), fN mN (fN)b (mN)cfN mN fN (mN)d fN (mN)e, wherein N represents any nucleotide, m represents a 2’-0Me modification, f represents a 2’-F modification, a is an integer of 1 or greater, each of b, c, d, and e are independently an integer of 2 or greater, and the sum of a, b, c, d, and e is not greater than 22. In some embodiments, a sense strand comprises fN mN fN (mN)gfN mN (fN)h (mN)i f (mN)j, wherein N represents any nucleotide, m represents a 2’-0Me modification, f represents a 2’-F modification, each of g, h, i, and j are independently an integer of 2 or greater, and the sum of g, h, i, and j is not greater than 24. In some embodiments, an antisense strand comprises mN fN (mN), fN mN (fN)b (mN)cfN mN fN (mN)d fN (mN)eand a sense strand comprises fN mN fN (mN)gfN mN (fN)h (mN)i fN (mN)j, wherein N represents any nucleotide, m represents a 2’-OMe modification, f represents a 2’-F modification, a is an integer of 1 or greater, each of b, c, d, e, g, h, i, and j are independently an integer of 2 or greater, the sum of a, b, c, d, and e is not greater than 22, and the sum of g, h, i, and j is not greater than 24.

[0008] In some embodiments, a sum of a, b, c, d, and e is not greater than 15. In some embodiments, b, d, and e are each independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, b, d, and e are each independently an integer selected from 2, 3, 4, or 5. In some embodiments, c is an integer of 3 or greater. In some embodiments, c is an integer selected from 3, 4, 5, 6, 7, or 8. In some embodiments, c is an integer selected from 3, 4, or 5. In some embodiments, b is 2. In some embodiments, c is 4. In some embodiments, d is 3. In some embodiments, e is 3. In some embodiments, a is an integer of 2 or greater. In some embodiments, a is an integer selected from 2, 3, 4, 5, 6, or 7. In some embodiments, a is an integer selected from 2, 3, 4, or 5. In some embodiments, a is 3. In some embodiments, a is 1.

[0009] In some embodiments, a sum of g, h, i, and j is not greater than 15. In some embodiments, g, h, i, and j are each independently an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, g, h, i, and j are each independently an integer selected from 2, 3, 4, 5, or 6. In some embodiments, g is 3. In some embodiments, h is 3. In some embodiments, i is 3. In some embodiments, j is 6.

[0010] In some embodiments, an antisense strand comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an antisense strand comprisesAttorney Docket No.: 2010794-3211mN fN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, a sense strand comprises fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN and a sense strand comprises fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN and a sense strand comprises f mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN.

[0011] In some embodiments, an antisense strand comprises mN fN (mN)afN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleotide, m represents a 2’-OMe modification, f represents a 2’-F modification, a is an integer of 1 or greater, each of b, c, and d are independently an integer of 2 or greater, and the sum of a, b, c, and d is not greater than 23. In some embodiments, a sense strand (mN)gfN mN (fN)h (mN)i, wherein N represents any nucleotide, m represents a 2’-OMe modification, f represents a 2’-F modification, each of g, h, i, and j are independently an integer of 2 or greater, and the sum of g, h, and i is not greater than 28. In some embodiments, an antisense strand comprises mN fN (mN)afN mN (fN)b (mN)cfN mN fN (mN)d and a sense strand comprises (mN). fN mN (fN)h (mN)i, wherein N represents any nucleotide, m represents a 2’-OMe modification, f represents a 2’-F modification, a is an integer of 1 or greater, each of b, c, d, g, h, and i are independently an integer of 2 or greater, the sum of a, b, c, and d is not greater than 23, and the sum of g, h, and i is not greater than 28.

[0012] In some embodiments, a sum of a, b, c, and d is not greater than 16. In some embodiments, b and d are each independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, b is an integer selected from 2, 3, 4, or 5. In some embodiments, d is an integer selected from 4, 5, 6, 7, or 8. In some embodiments, c is an integer of 3 or greater. In some embodiments, c is an integer selected from 3, 4, 5, 6, 7, or 8. In some embodiments, c is an integer selected from 3, 4, or 5. In some embodiments, b is 2. In some embodiments, c is 4. In some embodiments, d is 7. In some embodiments, a is an integer of 2 or greater. In some embodiments, a is an integer selected from 2, 3, 4, 5, 6, or 7. In some embodiments, a is an integer selected from 2, 3, 4, or 5. In some embodiments, a is 3.

[0013] In some embodiments, a sum of g, h, and i is not greater than 19. In some embodiments, g, h, and i are each independently an integer selected from 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. In some embodiments, g and h are each independently an integer selected from 2, 3, 4, 5, or 6. In some embodiments, i is an integer selected from 6, 7, 8, 9, 10, 11, or 12. In some embodiments, g is 6. In some embodiments, h is 3. In some embodiments, i is 10.

[0014] In some embodiments, an antisense strand comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, a sense strand comprises mN mN mN mN mN mN fN mN fN fN fN mN mN mN mN mN mN mN mN mN mN. In someAttorney Docket No.: 2010794-3211embodiments, an antisense strand comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN and a sense strand comprises mN mN mN mN mN mN fN mN fN fN fN mN mN mN mN mN mN mN mN mN mN.

[0015] In some embodiments, an oligonucleotide comprises one or more phosphodiester linkages. In some embodiments, an oligonucleotide comprises one or more modified internucleotidic linkages. In some embodiments, an oligonucleotide comprises one or more phosphorothioate internucleotidic linkages. In some embodiments, an antisense strand comprises one or more phosphodiester linkages. In some embodiments, an antisense strand comprises one or more modified internucleotidic linkages. In some embodiments, an antisense strand comprises one or more phosphorothioate internucleotidic linkages. In some embodiments, an antisense strand comprises at least 2, 3, or 4 phosphorothioate internucleotidic linkages. In some embodiments, an antisense strand comprises at least 2 phosphorothioate internucleotidic linkages at the 5’ end. In some embodiments, an antisense strand comprises at least 2 phosphorothioate internucleotidic linkages at the 3’ end. In some embodiments, an antisense strand comprises mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, a sense strand comprises one or more phosphodiester linkages. In some embodiments, a sense strand comprises one or more modified internucleotidic linkages. In some embodiments, a sense strand comprises one or more phosphorothioate internucleotidic linkages. In some embodiments, a sense strand comprises at least 2, 3, or 4 phosphorothioate internucleotidic linkages. In some embodiments, a sense strand comprises at least 2 phosphorothioate internucleotidic linkages at the 5 ’ end. In some embodiments, a sense strand comprises at least 2 phosphorothioate internucleotidic linkages at the 3’ end. In some embodiments, a sense strand comprises fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, a sense strand comprises mN * mN * mN mN mN mN fN mN fN fN fN mN mN mN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0016] In some embodiments, an oligonucleotide comprises one or more internucleotidic linkages that are independently chirally controlled. In some embodiments, an oligonucleotide comprises one or more phosphorothioate internucleotidic linkages that are independently chirally controlled. In some embodiments, an antisense strand comprises one or more internucleotidic linkages that are independently chirally controlled. In some embodiments, an antisense strand comprises one or more phosphorothioateAttorney Docket No.: 2010794-3211internucleotidic linkages that are independently chirally controlled. In some embodiments, a sense strand comprises one or more internucleotidic linkages that are independently chirally controlled. In some embodiments, a sense strand comprises one or more phosphorothioate internucleotidic linkages that are independently chirally controlled.

[0017] In some embodiments, an oligonucleotide comprises one or more modified nucleobases. In some embodiments, an antisense strand comprises one or more modified nucleobases. In some embodiments, a sense strand comprises one or more modified nucleobases.

[0018] In some embodiments, a sense strand and an antisense strand can hybridize. In some embodiments, a sense strand and an antisense strand can hybridize and form a duplex region. In some embodiments, a duplex region is at least 15, 16, 17, 18, 19. 20, or 21 nucleosides in length. In some embodiments, a duplex region is 15, 16, 17, 18, 19, 20, or 21 nucleosides in length.

[0019] In some embodiments, an oligonucleotide comprises a blunt end. In some embodiments, an oligonucleotide comprises an overhang. In some embodiments, an oligonucleotide comprises two overhangs. In some embodiments, an oligonucleotide comprises an overhang on an antisense strand. In some embodiments, an oligonucleotide comprises an overhang on a sense strand. In some embodiments, an oligonucleotide comprises an overhang on an antisense strand and an overhang on a sense strand. In some embodiments, an oligonucleotide comprises an overhang on a 3’ end of an antisense strand. In some embodiments, an oligonucleotide comprises an overhang on a 3’ end of a sense strand. In some embodiments, an overhang comprises at least 1, 2, or 3 nucleosides. In some embodiments, an overhang comprises 1, 2, or 3 nucleosides. In some embodiments, an overhang comprises 1 nucleoside. In some embodiments, an overhang comprises 2 nucleosides. In some embodiments, an overhang comprises 3 nucleosides.

[0020] In some embodiments, an oligonucleotide comprise a 5 ’-terminal group. In some embodiments, a 5 ’-terminal group is located on the 5’ end of an antisense strand. In some embodiments, a 5 ’-terminal group comprises or is 5 ’-vinyl phosphonate, 5’-methylphosphonate, 5’-C-methyl with phosphate, or 5 ’-phosphorothioate. In some embodiments, a 5 ’-vinyl phosphonate is 5’-(£)-vinyl phosphonate. In some embodiments, a 5’-C-methyl with phosphate is (S) -5 ’-C -methyl with phosphate. In some embodiments, a 5 ’-terminal group comprises or is 5 ’-vinyl phosphonate. In some embodiments, a 5 ’-terminal group comprises or is 5’-(E)-vinyl phosphonate. In some embodiments, a 5 ’-terminal group comprises or is 5’-methylphosphonate. In some embodiments, a 5’-terminal group comprises or is 5’-C-methyl with phosphate. In some embodiments, a 5’-terminal group comprises or is (S)-5’-C-methyl with phosphate. In some embodiments, a 5 ’-terminal group comprises or is 5 ’-phosphorothioate.

[0021] In some embodiments, an oligonucleotide comprises a modification pattern of an oligonucleotide in Table 1 (e.g., Table 1 A, Table IB, Table 1 C), Table 3, or Table 4. In some embodiments,Attorney Docket No.: 2010794-3211an oligonucleotide comprises a sense strand and an antisense strand, wherein the sense strand comprises a modification pattern of a sense strand in Table 1 (e.g., Table 1 A, Table IB, Table 1C), Table 3, or Table 4. In some embodiments, an oligonucleotide comprises a sense strand and an antisense strand, wherein the antisense strand comprises a modification pattern of an antisense strand in Table 1 (e.g., Table 1A, Table IB, Table 1C), Table 3, or Table 4. In some embodiments, an oligonucleotide comprises a sense strand and an antisense strand, wherein the sense strand comprises a modification pattern of a sense strand in Table 1 (e.g., Table 1 A, Table IB, Table 1C), Table 3, or Table 4 and the antisense strand comprises a modification pattern of an antisense strand in Table 1 (e.g., Table 1A, Table IB, Table 1C), Table 3, or Table 4. In some embodiments, an oligonucleotides comprises a sense strand and an antisense strand, wherein the sense strand comprises a modification pattern of a sense strand in a row in Table 1 (e.g., Table 1A, Table IB, Table 1C), Table 3, or Table 4 and the antisense strand comprises a modification pattern of an antisense strand in the same row in Table 1 (e.g., Table 1 A, Table IB, Table I C), Table 3, or Table 4.

[0022] In some embodiments, an antisense strand comprises a base sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to a target sequence of the same length in a target RNA. In some embodiments, an antisense strand has a base sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to a target sequence of the same length in a target RNA. In some embodiments, a sense strand comprises a base sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a target sequence of the same length in a target RNA. In some embodiments, a sense strand has a base sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a target sequence of the same length in a target RNA. In some embodiments, a target RNA comprises or is a mRNA.

[0023] In some embodiments, an oligonucleotide is conjugated to a targeting agent. In some embodiments, a targeting agent comprises an antibody an antibody, an antigen binding portion of an antibody, a carbohydrate, a fatty acid, a lipid, a nanoparticle, a nucleic acid, a peptide or polypeptide, a protein, a small molecule, or any combination thereof. In some embodiments, a targeting agent is conjugated to an oligonucleotide by way of a linker.

[0024] In some embodiments, an oligonucleotide is capable of providing knockdown of a target RNA. In some embodiments, an oligonucleotide comprises or is an RNAi oligonucleotide. In some embodiments, an oligonucleotide comprises or is an siRNA.

[0025] In some embodiments, an oligonucleotide is a pharmaceutically acceptable salt.

[0026] In some embodiments, the present disclosure provides compositions comprising an oligonucleotide, e.g., an oligonucleotide provided herein. In some embodiments, a composition comprises an oligonucleotide provided herein. In some embodiments, a composition comprises one or moreAttorney Docket No.: 2010794-3211pharmaceutically acceptable salts of an oligonucleotide provided herein. In some embodiments, a composition comprising an oligonucleotide provided herein is a liquid composition.

[0027] In some embodiments, a composition comprises an oligonucleotide provided herein and a delivery agent. In some embodiments, a delivery agent comprises an antibody, an antigen binding portion of an antibody, a carbohydrate, a fatty acid, a lipid, a nanoparticle, a nucleic acid, a peptide or polypeptide, a small molecule, or any combination thereof. In some embodiments, a delivery agent comprises a nanoparticle. In some embodiments, a nanoparticle comprises a lipid nanoparticle (LNP) or a protein vault. In some embodiments, a delivery agent comprises a liposome, a lipoplex, or an LNP. In some embodiments, a delivery agent encapsulates an oligonucleotide, e.g., an oligonucleotide provided herein.

[0028] In some embodiments, the present disclosure provides pharmaceutical compositions comprising an oligonucleotide, e.g., an oligonucleotide provided herein and a pharmaceutically acceptable carrier. In some embodiments, a pharmaceutical composition comprises an oligonucleotide provided herein and a pharmaceutically acceptable carrier. In some embodiments, a pharmaceutical composition comprises one or more pharmaceutically acceptable salts of an oligonucleotide provided herein and a pharmaceutically acceptable carrier. In some embodiments, a pharmaceutical composition comprising an oligonucleotide provided herein and a pharmaceutically acceptable carrier is a liquid composition. In some embodiments, a pharmaceutically acceptable carrier comprises a buffer. In some embodiments, a pharmaceutically acceptable carrier comprises a buffered saline.

[0029] In some embodiments, the present disclosure provides methods (e.g., for reducing level of a target RNA, polypeptide encoded by a target RNA, and / or activity of a protein encoded by a target RNA in a system) comprising administering or delivering to a system an oligonucleotide, composition, or pharmaceutical composition provided herein. In some embodiments, a method comprises administering or delivering to a system an oligonucleotide, composition, or pharmaceutical composition provided herein. In some embodiments, a method for reducing level of a target RNA in a system comprises administering or delivering to the system an oligonucleotide, composition, or pharmaceutical composition provided herein. In some embodiments, a method for reducing level of polypeptide encoded by a target RNA in a system comprises administering or delivering to the system an oligonucleotide, composition, or pharmaceutical composition provided herein. In some embodiments, a method for reducing level of activity of a protein in a system comprises administering or delivering to the system an oligonucleotide, composition, or pharmaceutical composition provided herein, wherein the protein is encoded by a target RNA in the system. In some embodiments, a target RNA comprises or is mRNA.

[0030] In some embodiments, a system comprises or expresses a target RNA, e.g., a target mRNA. In some embodiments, a system comprises or is a cell. In some embodiments, a system comprises or is a tissue. In some embodiments, a system comprises or is an organ. In some embodiments, a systemAttorney Docket No.: 2010794-3211comprises or is an organism. In some embodiments, a system comprises or is a mammal. In some embodiments, a system comprises or is a human. In some embodiments, a system comprises or is an adult. In some embodiments, a system comprises or is a human adult. In some embodiments, a system comprises or is a child. In some embodiments, a system comprises or is a human child.

[0031] In some embodiments, the present disclosure provides methods for preventing or treating a condition, disorder, or disease, comprising administering or delivering to a subject susceptible thereto or suffering therefrom a therapeutically effective amount of an oligonucleotide, composition, or pharmaceutical composition provided herein. In some embodiments, a method for preventing a condition, disorder, or disease comprises administering or delivering to a subject susceptible thereto a therapeutically effective amount of an oligonucleotide, composition, or pharmaceutical composition provided herein. In some embodiments, onset of a condition, disorder, or disease is delayed. In some embodiments, onset of a condition, disorder, or disease is prevented. In some embodiments, a method for treating a condition, disorder, or disease comprises administering or delivering to a subject suffering therefrom a therapeutically effective amount of an oligonucleotide, composition, or pharmaceutical composition provided herein.

[0032] In some embodiments, a subject is a mammal. In some embodiments, a subject is a human. In some embodiments, a subject is an adult. In some embodiments, a subject is a human adult. In some embodiments, a subject is a child. In some embodiments, a subject is a human child.

[0033] In some embodiments, an oligonucleotide is administered or delivered using a LNP. In some embodiments, an oligonucleotide is administered or delivered using a liposome. In some embodiments, an oligonucleotide is administered or delivered using an exosome.

[0034] In some embodiments, an oligonucleotide, composition, or pharmaceutical composition is administered or delivered parenterally. In some embodiments, an oligonucleotide, composition, or pharmaceutical composition is administered or delivered intramuscularly. In some embodiments, an oligonucleotide, composition, or pharmaceutical composition is administered or delivered intravenously. In some embodiments, an oligonucleotide, composition, or pharmaceutical composition is administered or delivered subcutaneously.

[0035] In some embodiments, the present disclosure provides uses of oligonucleotides, compositions, or pharmaceutical compositions provided herein. In some embodiments, the present disclosure provides a method for preparing an oligonucleotide, composition, or pharmaceutical composition provided herein.BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1. Schematic of certain oligonucleotide modification patterns (Initial, ST0, STI, ST2,Attorney Docket No.: 2010794-3211and ST3) provided herein. Initial modification pattern comprised a sense strand comprising mN * mN * fN mN fN mN fN mN fN mN fN mN fN mN fN mN fN * mN * fN and an antisense strand comprising mN * fN * mN fN mN fN mN fN mN fN mN fN mN fN mN fN mN fN mN * mN * mN, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and * represents a phosphorothioate internucleotidic linkage. STO modification pattern comprised a sense strand comprising mN * mN * fN mN fN mN fN mN fN mN fN mN fN mN fN mN fN mN fN * mN * fN and an antisense strand comprising mN * f * mN fN mN fN mN fN mN fN mN fN mN fN mN fN mN fN mN fN mN * mN * mN, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and * represents a phosphorothioate internucleotidic linkage. STI modification pattern comprised a sense strand comprising fN * mN * fN mN mN mN fN mN fN fN fN fN mN mN fN mN mN mN mN * mN * mN and an antisense strand comprising mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein N represents any nucleoside, m represents a 2 ’ -OMe modification, f represents a 2 ’ -F modification, and * represents a phosphorothioate internucleotidic linkage. ST2 modification pattern comprised a sense strand comprising fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN and an antisense strand comprising mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein N represents any nucleoside, m represents a 2’ -OMe modification, f represents a 2’-F modification, and * represents a phosphorothioate internucleotidic linkage. ST3 modification pattern comprised a sense strand comprising mN * mN * fN mN fN mN fN mN fN fN fN mN fN mN fN mN fN mN fN * mN * fN and an antisense strand comprising mN * fN * mN fN mN fN mN fN mN fN mN mN mN fN mN fN mN fN mN fN mN * mN * mN, wherein N represents any nucleoside, m represents a 2 ’-OMe modification, f represents a 2’-F modification, and * represents a phosphorothioate internucleotidic linkage.

[0037] Figure 2. Graph of exemplary data of knockdown of NtrkA in vitro. Oligonucleotides targeting NtrkA comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested. Plotted lines at 0.00256 nM represent, from top to bottom, data for Initial 1003 and ST2 1003. Error bars represent SEM.

[0038] Figure 3. Graph of exemplary data of knockdown of NtrkA in vitro. Oligonucleotides targeting NtrkA comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested. Plotted lines at 0.00256 nM represent, from top to bottom, data for Initial 1519 and ST2 1519; plotted lines at 40 nM represent, from top to bottom, data for ST2 1519 and Initial 1519. Error bars represent SEM.

[0039] Figure 4. Graph of exemplary data of knockdown of NtrkA in vitro. Oligonucleotides targeting NtrkA comprising the Initial modification pattern, the ST2 modification pattern, the STO modification pattern, or the ST3 modification pattern as depicted in Figure 1 were designed and tested. Plotted lines atAttorney Docket No.: 2010794-32110.3125 nM represent, from top to bottom, data for ST22011, Initial 2011 , STO 2011 , and ST32011; plotted lines at 20 nM represent, from top to bottom, data for ST2 2011, Initial 2011, ST3 2011, and STO 2011. Error bars represent SEM.

[0040] Figure 5. Graph of exemplary data of knockdown of NtrkA in vitro. Oligonucleotides targeting NtrkA comprising the Initial modification pattern, the ST2 modification pattern, the STO modification pattern, or the ST3 modification pattern as depicted in Figure 1 were designed and tested. Plotted lines at 0.3125 nM represent, from top to bottom, data for Initial 2473, ST32473, ST22473, and STO 2473; plotted lines at 2.5 nM represent, from top to bottom, data for ST3 2473, Initial 2473, ST2 2473, and STO 2473. Error bars represent SEM.

[0041] Figure 6. Schematic of certain oligonucleotide modification patterns (Initial, ST2-V) provided herein. Initial modification pattern comprised a sense strand comprising mN * mN * fN mN fN mN fN mN fN mN fN mN fN mN fN mN fN mN fN * mN * fN and an antisense strand comprising mN * fN * mN fN mN fN mN fN mN fN mN fN mN fN mN fN mN fN mN * fN * mN, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and * represents a phosphorothioate internucleotidic linkage. ST2-V modification pattern comprised a sense strand comprising fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN and an antisense strand comprising mN * fN * mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and * represents a phosphorothioate internucleotidic linkage.

[0042] Figure 7A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2641. The X-axis depicts concentration of tested oligonucleotide (RT2641) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0043] Figure 7B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2641VP. The X-axis depicts concentration of tested oligonucleotide (RT2641VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0044] Figure 8A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2647. The X-axis depicts concentration of tested oligonucleotide (RT2647) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.Attorney Docket No.: 2010794-3211

[0045] Figure 8B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2647VP. The X-axis depicts concentration of tested oligonucleotide (RT2647VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0046] Figure 9A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2655. The X-axis depicts concentration of tested oligonucleotide (RT2655) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0047] Figure 9B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2655VP. The X-axis depicts concentration of tested oligonucleotide (RT2655VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0048] Figure 10A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2657. The X-axis depicts concentration of tested oligonucleotide (RT2657) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0049] Figure 10B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2657VP. The X-axis depicts concentration of tested oligonucleotide (RT2657VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0050] Figure 11 A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2665. The X-axis depicts concentration of tested oligonucleotide (RT2665) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2. LEUTX). Error bars represent SEM.

[0051] Figure 1 IB. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2665VP. The X-axis depicts concentration of tested oligonucleotide (RT2665VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCRAttorney Docket No.: 2010794-3211(DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0052] Figure 12A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2677. The X-axis depicts concentration of tested oligonucleotide (RT2677) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0053] Figure 12B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2677VP. The X-axis depicts concentration of tested oligonucleotide (RT2677VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0054] Figure 13 A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2701. The X-axis depicts concentration of tested oligonucleotide (RT2701) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0055] Figure 13B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2701 VP. The X-axis depicts concentration of tested oligonucleotide (RT2701VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0056] Figure 14A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2702. The X-axis depicts concentration of tested oligonucleotide (RT2702) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0057] Figure 14B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2702VP. The X-axis depicts concentration of tested oligonucleotide (RT2702VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0058] Figure 15A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2703. The X-axis depicts concentration of tested oligonucleotideAttorney Docket No.: 2010794-3211(RT2703) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0059] Figure 15B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2703VP. The X-axis depicts concentration of tested oligonucleotide (RT2703VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0060] Figure 16A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2705. The X-axis depicts concentration of tested oligonucleotide (RT2705) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0061] Figure 16B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2705VP. The X-axis depicts concentration of tested oligonucleotide (RT2705VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0062] Figure 17A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2708. The X-axis depicts concentration of tested oligonucleotide (RT2708) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0063] Figure 17B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2708VP. The X-axis depicts concentration of tested oligonucleotide (RT2708VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0064] Figure 18 A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2721. The X-axis depicts concentration of tested oligonucleotide (RT2721) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0065] Figure 18B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalizedAttorney Docket No.: 2010794-3211FSHD myoblasts transfected with RT2721 VP. The X-axis depicts concentration of tested oligonucleotide (RT2721VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0066] Figure 19 A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2724. The X-axis depicts concentration of tested oligonucleotide (RT2724) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0067] Figure 19B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2724VP. The X-axis depicts concentration of tested oligonucleotide (RT2724VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0068] Figure 20A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2782. The X-axis depicts concentration of tested oligonucleotide (RT2782) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0069] Figure 20B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2782VP. The X-axis depicts concentration of tested oligonucleotide (RT2782VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0070] Figure 21 A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2789. The X-axis depicts concentration of tested oligonucleotide (RT2789) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0071] Figure 2 IB. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2789VP. The X-axis depicts concentration of tested oligonucleotide (RT2789VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0072] Figure 22A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulatedAttorney Docket No.: 2010794-3211mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2791. The X-axis depicts concentration of tested oligonucleotide (RT2791) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0073] Figure 22B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2791 VP. The X-axis depicts concentration of tested oligonucleotide (RT2791VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0074] Figure 23A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2792. The X-axis depicts concentration of tested oligonucleotide (RT2792) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0075] Figure 23B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2792VP. The X-axis depicts concentration of tested oligonucleotide (RT2792VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0076] Figure 24A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2796. The X-axis depicts concentration of tested oligonucleotide (RT2796) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2. LEUTX). Error bars represent SEM.

[0077] Figure 24B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2796VP. The X-axis depicts concentration of tested oligonucleotide (RT2796VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0078] Figure 25 A. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2928. The X-axis depicts concentration of tested oligonucleotide (RT2928) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.Attorney Docket No.: 2010794-3211

[0079] Figure 25B. Graph of exemplary data of knockdown of DUX4 mRNA and DUX4-regulated mRNAs (ZSCAN4, MBD3L2, LEUTX) in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with RT2928VP. The X-axis depicts concentration of tested oligonucleotide (RT2928VP) transfected; the Y-axis depicts percent remaining mRNA as measured by RT-digital PCR (DUX4) or RT-qPCR (ZSCAN4, MBD3L2, LEUTX). Error bars represent SEM.

[0080] Figure 26. Graph of exemplary data of FSHD composite gene expression in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts FSHD composite gene expression (which comprises ZSCAN4, MBD3L2, LEUTX) as a percentage of mock control. Error bars represent SEM.

[0081] Figure 27. Graphs of exemplary data of differentially expressed genes as determined by RNAseq from samples collected from FSHD myotubes (ABIC; left graph) or unaffected myotubes (UB1C; right graph). For each comparison, genes with decreased expression levels are represented by a column extending downward from 0 and genes with increased expression levels are represented by a column extending upward from 0. The X-axis displays comparison (e.g., ABIC vs. UBIC, RT2641 vs. vehicle) and indicates the cell type tested (e.g., ABIC myotube, UBIC myotube).

[0082] Figure 28. Graphs of exemplary data of measured expression levels of DUX4 as determined by RNAseq. Top graph displays results from FSHD myotubes (ABIC) and bottom graph displays results from unaffected myotubes (UBIC). TPM = # of transcripts per million total transcripts.

[0083] Figure 29. Graphs of exemplary data of measured expression levels of certain DUX4-regulated genes (ZSCAN4, LEUTX, MBD3L2) as determined by RNAseq. Graphs on left display results from FSHD myotubes (ABIC) and graphs on right display results from unaffected myotubes (UBIC). TPM = # of transcripts per million total transcripts.

[0084] Figure 30. Graphs of exemplary data of composite score of expression levels of various DUX4-regulated genes. Panel A = LEUTX, TRIM43, MBD3L2, and KHDC1L; Panel B = CCNA1, ZSCAN4, MBD3L2, KHDC1L, SLC34A2, and PRAMEF6; 41-Gene Panel = gene panel as described in van den Heuvel et al. Sci Rep. 2022;12(l):1426.

[0085] Figure 31. Graphs of exemplary data of off-target analysis for various oligonucleotides using RNAseq of samples from FSHD (ABIC) myotubes.

[0086] Figure 32. Graphs of exemplary data of off-target analysis for various oligonucleotides using RNAseq of samples from unaffected (UBIC) myotubes.

[0087] Figure 33A. Graph of exemplary data of knockdown of Lp(a) mRNA in vitro. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested. Plotted lines at 0.25 nM represent, from top to bottom, data for Lp(a)Attorney Docket No.: 2010794-32111271 Initial and Lp(a) 1271 ST2.

[0088] Figure 33B. Graph of exemplary data of knockdown of Lp(a) mRNA in vitro. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested. Plotted lines at 0.25 nM represent, from top to bottom, data for Lp(a) 2807 ST2 and Lp(a) 2807 Initial.

[0089] Figure 33C. Graph of exemplary data of knockdown of Lp(a) mRNA in vitro. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested. Plotted lines at 0.25 nM represent, from top to bottom, data for Lp(a) 3617 ST2 and Lp(a) 3617 Initial.

[0090] Figure 33D. Graph of exemplary data of knockdown of Lp(a) mRNA in vitro. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested. Plotted lines at 0.25 nM represent, from top to bottom, data for Lpia) 3635 ST2 and Lp(a) 3635 Initial.

[0091] Figure 33E. Graph of exemplary data of knockdown of Lp(a) mRNA in vitro. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested. Plotted lines at 0.25 nM represent, from top to bottom, data for Lp(a) 3725 Initial and Lp(a) 3725 ST2.

[0092] Figure 33F. Graph of exemplary data of knockdown of Lp(a) mRNA in vitro. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested. Plotted lines at 0.25 nM represent, from top to bottom, data for Lp(a) 4133 Initial and Lp(a) 4133 ST2.

[0093] Figure 33G. Graph of exemplary data of knockdown of Lp(a) mRNA in vitro. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested. Plotted lines at 0.25 nM represent, from top to bottom, data for Lp(a) 5447 ST2 and Lp(a) 5447 Initial; plotted lines at 16 nM represent, from top to bottom, data for Lp(a) 5447 Initial and Lp(a) 5447 ST2.

[0094] Figure 34A. Graph of exemplary data of in vitro knockdown of Lp(a) in Hep3B-hLPa cells. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and knockdown of Lp(a) mRNA at 24 hours and 48 hours post-transfection was assessed. For each time point, columns from left-to-right represent Lp(a) 1271 Initial (0.5 nM), Lp(a) 1271 Initial (10 nM), Lp(a) 1271 ST2 (0.5 nM), Lp(a) 1271 ST2 (10 nM). Inset dagger, double dagger, untiled circle, and unfiled rhombus also indicate columns corresponding to each treatment.

[0095] Figure 34B. Graph of exemplary data of in vitro knockdown of Lp(a) in Hep3B-hLPa cells. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modificationAttorney Docket No.: 2010794-3211pattern as depicted in Figure 1 were designed and knockdown of Lp(a) mRNA at 24 hours and 48 hours post-transfection was assessed. For each time point, columns from left-to-right represent Initial (0.5 nM), Lp(a) 3725 Initial (10 nM), Lp(a) 3725 ST2 (0.5 nM), Lp(a) 3725 ST2 (10 nM). Inset dagger, double dagger, unfiled circle, and unfiled rhombus also indicate columns corresponding to each treatment.

[0096] Figure 35 A. Graph of exemplary data of in vitro Ago2 loading with Lp(a) 1271 Initial or Lp(a) 1271 ST2 relative to miR-122 at either 24 hour or 48 hour time points post-transfection. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested for loading into Ago2. For each time point, columns from left-to-right represent Lp(a) 1271 Initial (0.5 nM), Lp(a) 1271 Initial (10 nM), Lp(a) 1271 ST2 (0.5 nM), Lp(a) 1271 ST2 (10 nM). Inset dagger, double dagger, unfiled circle, and unfiled rhombus also indicate columns corresponding to each treatment.

[0097] Figure 35B. Graph of exemplary data of in vitro Ago2 loading with Lp(a) 3725 Initial or Lp(a) 3725 ST2 relative to miR-122 at either 24 hours or 48 hours post-transfection. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested for loading into Ago2. For each time point, columns from left-to-right represent Lp(a) 3725 Initial (0.5 nM), Lp(a) 3725 Initial (10 nM), Lp(a) 3725 ST2 (0.5 nM), Lp(a) 3725 ST2 (10 nM). Inset dagger, double dagger, unfiled circle, and unfiled rhombus also indicate columns corresponding to each treatment.

[0098] Figure 36. Graph of exemplary data of in vitro knockdown of Lp(a) in Hep3B-hLPa cells. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested. For each oligonucleotide, columns from left-to-right represent data for 10 nM and 1 nM. Inset dagger, double dagger, unfiled circle, and unfiled rhombus also indicate columns corresponding to each concentration.

[0099] Figure 37. Graph of exemplary data of Ago2 loading with provided oligonucleotides relative to miR-122 at various time points post-transfection. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and tested. For each oligonucleotide, columns from left-to-right represent data for day 1, day 4, day 7, and day 14. Inset dagger and double dagger indicate columns corresponding to each timepoint.

[0100] Figure 38. Graph of exemplary data of human Lp(a) (hLPA) protein in serum. Mice were transduced with AAVs to introduce hLPA gene. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and administered. Levels of hLPA protein in serum were assessed prior to administration of AAV or oligonucleotides, post-AAV administration and prior to administration of oligonucleotides, and postadministration of oligonucleotides. For each time point, columns from left-to-right represent: GFP AAV +Attorney Docket No.: 2010794-3211PBS, hLPA AAV + PBS, hLPA AAV + Scrambled Control, hLPA AAV + GFP Control, hLPA AAV + RT2530, hLPA AAV + Lp(a) 3635 Initial GalNAc, hLPA AAV + Lp(a) 3725 ST2-P GalNAc, hLPA AAV + Lp(a) 5447 Initial GalNAc, hLPA AAV + Lp(a) 3617 Initial GalNAc, hLPA AAV + Lp(a) 1271 ST2 GalNAc (Prep 1), hLPA AAV + Lp(a) 1271 ST2-P GalNAc, and hLPA AAV + Lp(a) 1271 ST2 GalNAc (Prep 3). Five animals were used in each treatment group.

[0101] Figure 39. Graph of exemplary data of measured human Lp(a) (hLPA) mRNA in liver. Mice were transduced with AAVs to introduce hLPA gene. Oligonucleotides targeting Lp(a) comprising either the Initial modification pattern or the ST2 modification pattern as depicted in Figure 1 were designed and administered. Mice were sacrificed and hLPA mRNA levels in liver were assessed. Columns from left-to-right represent: GFP AAV + PBS, hLPA AAV + PBS, hLPA AAV + Scrambled Control, hLPA AAV + GFP Control, hLPA AAV + RT2530, hLPA AAV + Lp(a) 3635 Initial GalNAc, hLPA AAV + Lp(a) 3725 ST2-P GalNAc, hLPA AAV + Lp(a) 5447 Initial GalNAc, hLPA AAV + Lp(a) 3617 Initial GalNAc, hLPA AAV + Lp(a) 1271 ST2 GalNAc (Prep 1), hLPA AAV + Lp(a) 1271 ST2-P GalNAc, and hLPA AAV + Lp(a) 1271 ST2 GalNAc (Prep 2). Five animals were used in each treatment group.

[0102] Figure 40. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT2928, RT2657VP, RT2657.0, and RT2657.3. At 0.1 nM, from top to bottom, plotted lines represent data for RT2657VP, RT2657.0, RT2657.3, and RT2928. At 10 nM, from top to bottom, plotted lines represent data for RT2657VP, RT2657.3, RT2657.0, and RT2928.

[0103] Figure 41A. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT2702.5, RT2928, RT2702VP, RT2702.8, RT2702.0, and RT2702.13. At 0.333 nM, from top to bottom, plotted lines represent data for RT2702.5, RT2702.8, RT2702.13, RT2702VP, RT2702.0, and RT2928.

[0104] Figure 4 IB. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT2928, RT2702.il,Attorney Docket No.: 2010794-3211RT2702.12, RT2702VP, and RT2702.9. At 0.1 nM, from top to bottom, plotted lines represent data for RT2702.12, RT2928, RT2702VP, RT2702.9, and RT2702.il.

[0105] Figure 42A. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT2703.0, RT2928, RT2703.4, RT2703VP, and RT2703.5. At 1 nM, from top to bottom, plotted lines represent data for RT2703.5, RT2703VP, RT2703.4, RT2703.0, and RT2928.

[0106] Figure 42B. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT2703.7, RT2703VP, and RT2928. At 0.1 nM, from top to bottom, plotted lines represent data for RT2703.7, RT2928, and RT2703VP. At 10 nM, from top to bottom, plotted lines represent data for RT2703VP, RT2703.7, and RT2928.

[0107] Figure 43 A. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT2728.0, RT2928, RT2728VP, RT2728.4, and RT2728.13. At 0.1 nM, from top to bottom, plotted lines represent data for RT2728VP, RT2928, RT2728.0, RT2728.13, and RT2728.4.

[0108] Figure 43B. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT2728.16, RT2728.9, RT2928, RT2728.10, RT2728.11, RT2728VP, and RT2728.17. At 1 nM, from top to bottom, plotted lines represent data for RT2728.11, RT2728.16, RT2728.10, RT2928, RT2728.17, RT2728VP, and RT2728.9.

[0109] Figure 44. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depictsAttorney Docket No.: 2010794-3211percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT2791VP, RT2928, RT2791.5, RT2791.1, RT2791.4, RT2791.0, RT2791.6, RT2791.3, RT2791.7, RT2791.2, and RT2791.8.

[0110] Figure 45 A. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT 2792.7, RT2792VP, RT2928, RT2792.8, RT2792.3, RT2792.14, RT2792.5, RT2792.2, RT2792.4, RT2792.14, RT2792.0, and RT2792.15.

[0111] Figure 45B. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT2928, RT2792VP, RT2792.16, RT2792.17, RT2792.9, RT2792.10, and RT2792.il. At 0.1 nM, from top to bottom, plotted lines represent data for RT2928, RT2792.16, RT2792VP, RT2792.17, RT2792.10, RT2792.9, and RT2792.il.

[0112] Figure 46A. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT2789.1, RT2789.7, RT2789.6, RT2789.5, RT2789.0, RT2928, RT2789.2, and RT2789VP.

[0113] Figure 46B. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicated oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT2789VP, RT27928, and RT2789.8. At 0.1 nM, from top to bottom, plotted lines represent data for RT2928, RT2789VP, and RT2789.8. At 1 nM, from top to bottom, plotted lines represent data for RT2789VP, RT2928, and RT2789.8.

[0114] Figure 47. Graph of exemplary data of knockdown of DUX4-associated mRNA in myotubes differentiated from patient-derived immortalized FSHD myoblasts transfected with indicatedAttorney Docket No.: 2010794-3211oligonucleotides. The X-axis depicts concentration of tested oligonucleotide transfected; the Y-axis depicts percent composite mRNA (ZSCAN4, MBD3L2, LEUTX) remaining as measured by RT-qPCR. Error bars represent SEM. At 0.001 nM, from top to bottom, plotted lines represent data for RT2928, RT2809VP, RT2809.0, RT2809.4, RT2809.5, and RT2809.3.DEFINITIONS

[0115] As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75thEd. Additionally, general principles of organic chemistry are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999, and “March’s Advanced Organic Chemistry”, 5thEd., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001.

[0116] As used herein in the present disclosure, unless otherwise clear from 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 used with “not limited to” or not), and “include” (whether used with “not limited to” or not) may be understood to encompass itemized components or steps whether presented by themselves or together 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 permit standard variation as would be understood by those of ordinary skill in the art; and (vi) where ranges are provided, endpoints are included.

[0117] Unless otherwise specified, description of oligonucleotides and elements thereof (e.g., base sequence, sugar modifications, internucleotidic linkages, linkage phosphorus stereochemistry, patterns thereof, etc.) is from 5’ to 3’. As those skilled in the art will appreciate, in some embodiments, oligonucleotides may be provided and / or utilized as various forms, e.g., salt forms, particularly pharmaceutically acceptable salt forms, e.g., sodium salts. As those skilled in the art will also appreciate, in some embodiments, individual oligonucleotides within a composition may be considered to be of the same constitution and / or structure even though, within such composition (e.g., a liquid composition), particular such oligonucleotides might be in different forms, e.g., salt form(s) (and may be dissolved and the oligonucleotide chain may exist as an anion form when, e.g., in a liquid composition) at a particular moment in time. For example, those skilled in the art will appreciate that, at a given pH, individual internucleotidic linkages along an oligonucleotide chain may be in an acid (H) form, or in one of a plurality of possible salt forms (e.g., a sodium salt, or a salt of a different cation, depending on which ions might be present in the preparation or composition), and will understand that, so long as their acid forms (e.g., replacing all cations, if any, with H+) are of the same constitution and / or structure, such individualAttorney Docket No.: 2010794-3211oligonucleotides may properly be considered to be of the same constitution and / or structure.

[0118] About: As used herein, the term “about”, when used herein in reference to a value, refers to a value that is similar, in context to the referenced value. In general, those skilled in the art, familiar with the context, will appreciate the relevant degree of variance encompassed by “about” in that context. For example, in some embodiments, the term “about” may encompass a range of values that within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value.

[0119] Alkyl : As used herein, the term “alkyl” is given its ordinary meaning in the art and may 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, alkyl has 1-100 carbon atoms. In certain embodiments, a straight chain or branched chain alkyl has about 1-20 carbon atoms in its backbone (e.g., C1-C20 for straight chain, C2-C20 for branched chain), and alternatively, about 1-10. In some embodiments, cycloalkyl rings have from about 3-10 carbon atoms in their ring structure where such rings are monocyclic, bicyclic, or polycyclic, and alternatively about 5, 6 or 7 carbons in the ring structure. In some embodiments, an alkyl group may be a lower alkyl group, wherein a lower alkyl group comprises 1-4 carbon atoms (e.g., C1-C4 for straight chain lower alkyls).

[0120] 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 some embodiments, the non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, cattle, a primate and / or a pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish and / or worms. In some embodiments, an animal may be a transgenic animal, a genetically engineered animal and / or a clone.

[0121] Characteristic portion: As used herein, the term “characteristic portion”, in the broadest sense, refers to a portion of a substance whose presence (or absence) correlates with presence (or absence) of a particular feature, attribute, or activity of the substance. In some embodiments, a characteristic portion of a substance is a portion that is found in the substance and in related substances that share the particular feature, attribute or activity, but not in those that do not share the particular feature, 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 nucleic acid is one that contains a number of, in some embodiments, a continuous stretch of, nucleobases that are characteristic of that nucleic acid.

[0122] Heteroatom: The term “heteroatom”, as used herein, means an atom that is not carbon or hydrogen. In some embodiments, a heteroatom is boron, oxygen, sulfur, nitrogen, phosphorus, or siliconAttorney Docket No.: 2010794-3211(including oxidized forms of nitrogen, sulfur, phosphorus, or silicon; charged forms of nitrogen (e.g., quaternized forms, forms as in iminium groups, etc.), phosphorus, sulfur, oxygen; etc.). In some embodiments, a heteroatom is silicon, phosphorus, oxygen, sulfur or nitrogen. In some embodiments, a heteroatom is silicon, oxygen, sulfur or nitrogen. In some embodiments, a heteroatom is oxygen, sulfur or nitrogen.

[0123] Identity: As used herein, the term “identity” refers to the overall relatedness between polymeric molecules, e.g., between nucleic acid molecules (e.g., oligonucleotides, DNA, RNA, etc.) and / or between polypeptide molecules. In some embodiments, polymeric molecules are considered to be “substantially identical” to one another 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 the percent identity of two nucleic acid or polypeptide sequences, for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second sequences for optimal alignment and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of a sequence aligned for comparison purposes 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 a reference sequence. The nucleotides at corresponding positions are then compared. When 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, then the molecules are identical at that position. The percent identity between the 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, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished 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), which has been incorporated into the ALIGN program (version 2.0). In some exemplary embodiments, nucleic acid sequence comparisons made with the ALIGN program use a PAM 120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix.

[0124] Internucleotidic linkage: As used herein, the phrase “internucleotidic linkage” refers generally to a linkage linking nucleoside units of an oligonucleotide or a nucleic acid. In some embodiments, an internucleotidic linkage is a phosphodiester linkage, as extensively found in naturally occurring DNA and RNA molecules (natural phosphate linkage (-OP(=O)(OH)O-), which as appreciated by those skilled in the art may exist as a salt form). In some embodiments, an internucleotidic linkage is a modified internucleotidic linkage (not a natural phosphate linkage). In some embodiments, an internucleotidicAttorney Docket No.: 2010794-3211linkage is a “modified internucleotidic linkage” wherein at least one oxygen atom or -OH of a phosphodiester linkage is replaced by a different organic or inorganic moiety. In some embodiments, such an organic or inorganic moiety is selected from =S, =Se, =NR’, -SR’, -SeR’, -N(R’)2, B(R’) , -S-, -Se-, and -N(R’)-, wherein each R’ is independently -H or an optionally substituted group selected from Ci-io aliphatic, Ce-i4 aryl, Ci-io heteroaliphatic having 1-5 heteroatoms, 5-10 membered heteroaryl having 1-5 heteroatoms and 3-10 membered heterocyclyl having 1-4 heteroatoms, or two or more R’ groups are taken together with their intervening atoms to from an optionally substituted 3-10 membered ring having 0-5 heteroatoms in addition to the intervening atoms. In some embodiments, a modified internucleotidic linkage is a phosphorothioate linkage. It is understood by a person of ordinary skill in the art that an internucleotidic linkage may exist as an anion or cation at a given pH due to the existence of acid or base moieties in the linkage.

[0125] In vitro: As used herein, the term “in vitro” refers to events that occur in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, etc., rather than within an organism (e.g., animal, plant and / or microbe).

[0126] In vivo: As used herein, the term “in vivo” refers to events that occur within an organism (e.g., animal, plant and / or microbe).

[0127] Linkage phosphorus: As used herein, the phrase “linkage phosphorus” is used to indicate that the particular phosphorus atom being referred to is the phosphorus atom present in the internucleotidic linkage, which phosphorus atom corresponds to the phosphorus atom of a phosphodiester internucleotidic linkage as occurs in naturally occurring DNA and RNA. In some embodiments, a linkage phosphorus atom is in a modified internucleotidic linkage, wherein each oxygen atom of a phosphodiester linkage is optionally and independently replaced by an organic or inorganic moiety. In some embodiments, a linkage phosphorus atom is chiral (e.g., as in phosphorothioate internucleotidic linkages). In some embodiments, a linkage phosphorus atom is achiral (e.g., as in natural phosphate linkages).

[0128] Modified nucleobase: The terms “modified nucleobase”, “modified base” and the like refer to a chemical moiety which is chemically distinct from a nucleobase, but which is capable of performing at least one function of a nucleobase. In some embodiments, a modified nucleobase is a nucleobase which comprises a modification. In some embodiments, a modified nucleobase is capable of at least one function of a nucleobase, e.g., forming a moiety in a polymer capable of base-pairing to a nucleic acid comprising an at least complementary sequence of bases. In some embodiments, a modified nucleobase is 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 oligonucleotides refer to a nucleobase that is not A, T, C, G or U.

[0129] Modified nucleoside: The term “modified nucleoside” refers to a moiety derived from or chemically similar to a natural nucleoside, but which comprises a chemical modification whichAttorney Docket No.: 2010794-3211differentiates it from a natural nucleoside. Non-limiting examples of modified nucleosides include those which comprise a modification at a base and / or a sugar. Non-limiting examples of modified nucleosides include those with a 2’ modification at a sugar. Non-limiting examples of modified nucleosides also include abasic nucleosides (which lack a nucleobase). In some embodiments, a modified nucleoside is capable of at least one function of a nucleoside, e.g., forming a moiety in a polymer capable of base-pairing to a nucleic acid comprising an at least complementary sequence of bases.

[0130] Modified nucleotide: The term “modified nucleotide” includes any chemical moiety which differs structurally from a natural nucleotide but is capable of performing at least one function of a natural nucleotide. In some embodiments, a modified nucleotide comprises a modification at a sugar, base and / or internucleotidic linkage. In some embodiments, a modified nucleotide comprises a modified sugar, modified nucleobase and / or modified internucleotidic linkage. In some embodiments, a modified nucleotide is capable of at least one function of a nucleotide, e.g., forming a subunit in a polymer capable of base-pairing to a nucleic acid comprising an at least complementary sequence of bases.

[0131] Modified sugar: The term “modified sugar” refers to a moiety that can replace a sugar. A modified sugar mimics the spatial arrangement, electronic properties, or some other physicochemical property of a sugar. In some embodiments, as described in the present disclosure, a modified sugar is substituted ribose or deoxyribose. In some embodiments, a modified sugar comprises a 2’ -modification. Examples of useful 2’ -modification are widely utilized in the art and described herein. In some embodiments, a 2 ’-modification is 2’-F. In some embodiments, a 2 ’-modification is 2’-OR, wherein R is optionally substituted Ci-io aliphatic. In some embodiments, a 2 ’-modification is 2’-OMe (2’-O-methyl). In some embodiments, a 2’ -modification is 2’-M0E (2’-O-methoxyethyl). In some embodiments, a modified sugar is a bicyclic sugar (e.g., a sugar used in LNA, BNA, etc.). In some embodiments, in the context of oligonucleotides, a modified sugar is a sugar that is not ribose or deoxyribose as typically found in natural RNA or DNA.

[0132] Nucleic acid: The term “nucleic acid”, as used herein, includes any nucleotides 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 a combination thereof. These terms refer to the primary structure of the molecules and, thus, include double- and single-stranded DNA, and double-and single-stranded RNA. These terms include, as equivalents, analogs of either RNA or DNA comprising modified nucleotides and / or modified polynucleotides, such as, though not limited to, methylated, protected and / or capped nucleotides or polynucleotides. The terms encompass poly- or oligo-ribonucleotides (RNA) and poly- or oligo-deoxyribonucleotides (DNA); RNA or DNA derived from N-glycosides or C-glycosides of nucleobases and / or modified nucleobases; nucleic acids derived from sugars and / or modified sugars; and nucleic acids derived from phosphate bridges and / or modified internucleotidic linkages. The termAttorney Docket No.: 2010794-3211encompasses nucleic acids containing any combinations of nucleobases, modified nucleobases, sugars, modified sugars, phosphate bridges or modified internucleotidic linkages. In some embodiments, a “nucleic acid” is or comprises ribonucleic acids (RNAs), deoxyribonucleic acids (DNAs), threose nucleic acids (TNAs), glycol nucleic acids (GNAs), peptide nucleic acids (PNAs), locked nucleic acids (LNAs, including LNA having a p-D-ribo configuration. a-LNA having an cc-L-ribo configuration (a diastereomer of LNA), 2’-amino-LNA having a 2’-amino functionalization, and 2’-amino-a-LNA having a 2’-amino functionalization), or a combination thereof. In some embodiments, a nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 1 10, 120, 130, 140, 150, 160, 170, 180, 190, 20, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000 or more residues long. In some embodiments, a nucleic acid is partly or wholly single stranded; in some embodiments, a nucleic acid is partly or wholly double stranded. In some embodiments a nucleic acid has a nucleotide sequence comprising at least one element that encodes, or is the complement of a sequence that encodes, a functional gene product such as a RNA, polypeptide, or protein.

[0133] Nucleobase: The term “nucleobase” refers to the parts of nucleic acids that are involved in the hydrogen-bonding that binds 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, a naturally-occurring nucleobases are modified adenine, guanine, uracil, cytosine, or thymine. In some embodiments, a naturally-occurring nucleobases are methylated adenine, guanine, uracil, cytosine, or thymine. In some embodiments, a nucleobase comprises a heteroaryl ring wherein a ring atom is nitrogen, and when in a nucleoside, the nitrogen is bonded to a sugar moiety. In some embodiments, a nucleobase comprises a heterocyclic ring wherein a ring atom is nitrogen, and when in a nucleoside, the nitrogen is bonded to a sugar moiety. In some embodiments, a nucleobase is a “modified nucleobase,” a nucleobase other than adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T). In some embodiments, a modified nucleobase is substituted A, T, C, G or U. In some embodiments, a modified nucleobase is a substituted tautomer of A, T, C, G, or U. In some embodiments, a modified nucleobases is methylated adenine, guanine, uracil, cytosine, or thymine. In some embodiments, a modified nucleobase mimics the spatial arrangement, electronic properties, or some other physicochemical property of the nucleobase and retains the property of hydrogen-bonding that binds one nucleic acid strand to another in a sequence specific manner. In some embodiments, a modified nucleobase can pair with all of the five naturally occurring bases (uracil, thymine, adenine, cytosine, or guanine) without substantially affecting the melting behavior, recognition by intracellular enzymes or activity of the oligonucleotide duplex. As used herein, the term “nucleobase” also encompasses structural analogs used in lieu of natural or naturally-occurring nucleotides, such as modified nucleobases andAttorney Docket No.: 2010794-3211nucleobase analogs. In some embodiments, a nucleobase is optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, a “nucleobase” refers to a nucleobase unit in an oligonucleotide or a nucleic acid (e.g., A, T, C, G or U as in an oligonucleotide or a nucleic acid).

[0134] Nucleoside: The term “nucleoside” refers to a moiety wherein a nucleobase or a modified nucleobase is covalently bound to a sugar or a modified sugar. In some embodiments, a nucleoside is a natural nucleoside, e.g., adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, or deoxycytidine. In some embodiments, a nucleoside is a modified nucleoside, e.g., a substituted natural nucleoside selected from adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, and deoxycytidine. In some embodiments, a nucleoside is a modified nucleoside, e.g., a substituted tautomer of a natural nucleoside selected from adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, and deoxycytidine. In some embodiments, a “nucleoside” refers to a nucleoside unit in an oligonucleotide or a nucleic acid.

[0135] Nucleotide: The term “nucleotide” as used herein refers to a monomeric unit of a polynucleotide that consists of a nucleobase, a sugar, and one or more internucleotidic linkages (e.g., phosphate linkages in natural DNA and RNA). The naturally occurring bases [guanine, (G), adenine, (A), cytosine, (C), thymine, (T), and uracil (U)] are derivatives of purine or pyrimidine, though it should be understood that naturally and non-naturally occurring base analogs are also included. The naturally occurring sugar is the pentose (five-carbon sugar) deoxyribose (which forms DNA) or ribose (which forms RNA), though it should be understood that naturally and non-naturally occurring sugar analogs are also included. Nucleotides are linked via internucleotidic linkages to form nucleic acids, or polynucleotides. Many internucleotidic linkages are known in the art (such as, though not limited to, phosphate, phosphorothioates, boranophosphates and the like). Artificial nucleic acids include PNAs (peptide nucleic acids), phosphotriesters, phosphorothionates, H-phosphonates, phosphoramidates, boranophosphates, methylphosphonates, phosphonoacetates, thiophosphonoacetates and other variants of the phosphate backbone of native nucleic acids, such as those described herein. In some embodiments, a natural nucleotide comprises a naturally occurring base, sugar and internucleotidic linkage. As used herein, the term “nucleotide” also encompasses structural analogs used in lieu of natural or naturally-occurring nucleotides, such as modified nucleotides and nucleotide analogs. In some embodiments, a “nucleotide” refers to a nucleotide unit in an oligonucleotide or a nucleic acid.

[0136] Oligonucleotide: The term “oligonucleotide” refers to a polymer or oligomer of nucleotides and may contain any combination of natural and non-natural nucleobases, sugars, and internucleotidic linkages. Oligonucleotides can be single-stranded or double-stranded. A single-stranded oligonucleotide can have double-stranded regions (formed by two portions of the single-stranded oligonucleotide) and aAttorney Docket No.: 2010794-3211double-stranded oligonucleotide, which comprises two oligonucleotide chains, can have single-stranded regions for example, at regions where the two oligonucleotide chains are not complementary to each other. Example oligonucleotides include, but are not limited to structural genes, genes including control and termination regions, self-replicating systems such as viral or plasmid DNA, single-stranded and doublestranded RNAi agents and other RNA interference reagents (RNAi agents or iRNA agents), shRNA, antisense oligonucleotides, ribozymes, microRNAs, microRNA mimics, supermirs, aptamers, antimirs, antagomirs, U1 adaptors, triplex-forming oligonucleotides, G-quadruplex oligonucleotides, RNA activators, immuno-stimulatory oligonucleotides, and decoy oligonucleotides.

[0137] One or more: As used herein, in some embodiments, “one or more” is 1-200, 1-150, 1-100, 1-90, 1-80, 1-70, 1-60, 1-50, 1-40, 1-30, or 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 one. In some embodiments, “one or more” is two. In some embodiments, “one or more” is three. In some embodiments, “one or more” is four. In some embodiments, “one or more” is five. In some embodiments, “one or more” is six. In some embodiments, “one or more” is seven. In some embodiments, “one or more” is eight. In some embodiments, “one or more” is nine. In some embodiments, “one or more” is ten. In some embodiments, “one or more” is at least one. In some embodiments, “one or more” is at least two. In some embodiments, “one or more” is at least three. In some embodiments, “one or more” is at least four. In some embodiments, “one or more” is at least five. In some embodiments, “one or more” is at least six. In some embodiments, “one or more” is at least seven. In some embodiments, “one or more” is at least eight. In some embodiments, “one or more” is at least nine. In some embodiments, “one or more” is at least ten.

[0138] Optionally substituted: As described herein, compounds, e.g., oligonucleotides, of the disclosure may contain optionally substituted and / or substituted moieties. In general, the term “substituted,” whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated 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 substituent may be either the same or different at every position. In some embodiments, an optionally substituted group is unsubstituted. Combinations of substituents envisioned by this disclosure are preferably those that result in the formation of stable or chemically feasible compounds. The term “stable,” as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein. Certain substituents are described below.Attorney Docket No.: 2010794-3211

[0139] Pharmaceutically acceptable: As used herein, the phrase “pharmaceutically acceptable” refers to those compounds, materials, compositions and / or dosage forms which 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.

[0140] Pharmaceutically acceptable carrier: As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion of the body, to another organ, or portion of the body. 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 can serve as pharmaceutically-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 carboxymethyl cellulose, ethyl cellulose 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; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; pH buffered solutions; polyesters, polycarbonates and / or polyanhydrides; and other non-toxic compatible substances employed in pharmaceutical formulations.

[0141] Pharmaceutically acceptable salt: The term “pharmaceutically acceptable salt”, as used herein, refers to salts of such compounds that are appropriate for use in pharmaceutical contexts, i.e., salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge, et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). In some embodiments, pharmaceutically acceptable salt include, but are not limited to, nontoxic acid addition salts, which are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with 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, pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate,Attorney Docket No.: 2010794-3211hemisulfate, heptanoate, hexanoate, hydroiodide, 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 salts, and the like. In some embodiments, a provided compound comprises one or more acidic groups, e.g., an oligonucleotide, and a pharmaceutically acceptable salt is an alkali, alkaline earth metal, or ammonium (e.g., an ammonium salt of N(R)s, wherein each R is independently defined and described in the present disclosure) salt. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. In some embodiments, a pharmaceutically acceptable salt is a sodium salt. In some embodiments, a pharmaceutically acceptable salt is a potassium salt. In some embodiments, a pharmaceutically acceptable salt is a calcium salt. In some embodiments, pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and aryl sulfonate. In some embodiments, a provided compound comprises more than one acid groups, for example, an oligonucleotide may comprise two or more acidic groups (e.g., in natural phosphate linkages and / or modified internucleotidic linkages). In some embodiments, a pharmaceutically acceptable salt, or generally a salt, of such a compound comprises two or more cations, which can be the same or different. In some embodiments, in a pharmaceutically acceptable salt (or generally, a salt), all ionizable hydrogen (e.g., in an aqueous solution with a pKa no more than about 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2; in some embodiments, no more than about 7; in some embodiments, no more than about 6; in some embodiments, no more than about 5; in some embodiments, no more than about 4; in some embodiments, no more than about 3) in the acidic groups are replaced with cations. In some embodiments, each phosphorothioate and phosphate group independently exists in its salt form (e.g., if sodium salt, -O-P(O)(SNa)-O- and -O-P(O)(ONa)-O-, respectively). In some embodiments, each phosphorothioate and phosphate internucleotidic linkage independently exists in its salt form (e.g., if sodium salt, -O-P(O)(SNa)-O- and -O-P(O)(ONa)-O-, respectively). In some embodiments, a pharmaceutically acceptable salt is a sodium salt of an oligonucleotide. In some embodiments, a pharmaceutically acceptable salt is a sodium salt of an oligonucleotide, wherein each acidic phosphate and modified phosphate group (e.g., phosphorothioate, phosphate, etc.), if any, exists as a salt form (all sodium salt).

[0142] Pharmaceutical composition: As used herein, the term “pharmaceutical composition” refers to an active agent, formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, an active agent is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effectAttorney Docket No.: 2010794-3211when administered to a relevant population. In some embodiments, pharmaceutical compositions may be specially formulated for administration in solid or liquid form, including those adapted for the following: oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin, lungs, or oral cavity; intravaginally or intrarectally, for example, as a pessary, cream, or foam; sublingually; ocularly; transdermally; or nasally, pulmonary, and to other mucosal surfaces.

[0143] Polypeptide: As used herein, the term “polypeptide” refers to a polymer of at least three amino acids. Those of ordinary skill in the art will appreciate that the term “polypeptide” is intended to be sufficiently general as to encompass not only polypeptides having a complete sequence recited herein, but also to encompass polypeptides that represent functional fragments (e.g., fragments retaining at least one activity) of such complete polypeptides. Moreover, those of ordinary skill in the art understand that protein sequences generally tolerate some substitution without destroying activity. Thus, any polypeptide that retains activity and shares at least about 30% to about 40% overall sequence identity, often greater than about 50%, 60%, 70%, or 80%, and further usually including at least one region of much higher identity, often greater than 90% or even 95%, 96%, 97%, 98%, 99%, or greater in one or more highly conserved regions, usually encompassing at least 3 amino acids to 4 amino acids and often up to 20 amino acids or more, with another polypeptide of the same class, is encompassed within the term “polypeptide” as used herein. Polypeptides may contain L-amino acids, D-amino acids, or both. Polypeptides may contain any of a variety of amino acid modifications or analogs known in the art. Useful modifications include, e.g., terminal acetylation, amidation, and / or methylation. In some embodiments, polypeptides comprise natural amino acids, non-natural amino acids, synthetic amino acids, or combinations thereof. The term “peptide” is generally used to refer to a polypeptide having a length of less than about 100 amino acids, less than about 50 amino acids, less than 20 amino acids, or less than 10 amino acids.

[0144] Subject: As used herein, the term “subject” or “test subject” refers to any organism to which a compound or composition is administered in accordance with the present disclosure e.g., for experimental, diagnostic, prophylactic and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans; insects; worms: etc.) and plants. In some embodiments, a subject is a human. In some embodiments, a subject may be suffering from and / or susceptible to a disease, disorder and / or condition.

[0145] Sugar: The term “sugar” refers to a monosaccharide or polysaccharide in closed and / or open form. In some embodiments, sugars are monosaccharides. In some embodiments, sugars areAttorney Docket No.: 2010794-3211polysaccharides. Sugars include, but are not limited to, ribose, deoxyribose, pentofuranose, pentopyranose, and hexopyranose moieties. As used herein, the term “sugar” also encompasses structural analogs used in lieu of conventional sugar molecules, such as glycol, polymer of which forms the backbone of the nucleic acid analog, glycol nucleic acid (“GNA”), morpholine, polymer of which forms the backbone of the nucleic acid analog, morpholino, etc. As used herein, the term “sugar” also encompasses structural analogs used in lieu of natural or naturally-occurring nucleotides, such as modified sugars and nucleotide sugars. In some embodiments, a sugar is a RNA or DNA sugar (ribose or deoxyribose). In some embodiments, a 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 may provide one or more desired properties, activities, etc. In some embodiments, a sugar is optionally substituted ribose or deoxyribose. In some embodiments, a “sugar” refers to a sugar unit in an oligonucleotide or a nucleic acid.

[0146] Susceptible to: An individual who is “susceptible to” a disease, disorder and / or condition is one who has a higher risk of developing the disease, disorder and / or condition than does a member of the general public. In some embodiments, an individual who is susceptible to a disease, disorder and / or condition is predisposed to have that 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.

[0147] Therapeutically effective amount: As used herein, the term “therapeutically effective amount” means an amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that elicits a desired biological response when administered as part of a therapeutic regimen. In some embodiments, a therapeutically effective amount of a substance is an amount that is sufficient, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, to treat, diagnose, prevent, and / or delay the onset of the disease, disorder, and / or condition. As will be appreciated by those of ordinary skill in this art, the effective amount of a substance may vary depending on such factors as the desired biological endpoint, the substance to be delivered, the target cell or tissue, etc. For example, the effective amount of compound in a formulation to treat a disease, disorder, and / or condition is the amount that alleviates, ameliorates, relieves, inhibits, prevents, delays onset of, reduces severity of and / or reducesAttorney Docket No.: 2010794-3211incidence of one or more symptoms or features 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.

[0148] Treat: As used herein, the term “treat,” “treatment,” or “treating” refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition. Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and / or condition. In some embodiments, treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and / or condition, for example for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and / or condition.

[0149] Vector: As used herein, the term “vector” refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked, e.g., into a cell (e.g., a host cell described herein), tissue, and / or organism. By way of non-limiting example, one type of vector is a “viral vector”, in which additional DNA segments may be ligated into a viral genome. Another type of vector is a “plasmid,” which refers to a circular double stranded DNA loop into which additional DNA segments may be ligated. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to as “expression vectors.”

[0150] Wild-type: As used herein, the term “wild-type” has its art-understood meaning that refers to an entity having a structure and / or activity as found in nature in a “normal” (as contrasted with mutant, diseased, altered, etc.) state or context. Those of ordinary skill in the art will appreciate that wild type genes and polypeptides often exist in multiple different forms (e.g., alleles).DETAILED DESCRIPTION OF CERTAIN EMBODIMENTSBase Sequences

[0151] Among other things, the present disclosure provides various oligonucleotides comprising various base sequences so that the oligonucleotides can target specific nucleic acids, e.g., specific target transcripts and affect for one or more biological functions. In some embodiments, oligonucleotides specifically target target nucleic acids. In some embodiments, oligonucleotides specifically target target transcripts. In some embodiments, a base sequence of a provided oligonucleotide is or comprises a sequence that is identical, complementary, or capable of hybridizing to a portion (a “target portion”) of aAttorney Docket No.: 2010794-3211target nucleic acid.

[0152] In some embodiments, a target portion comprises a portion of an exon and / or a portion of an intron. Exons and introns typically alternate, e.g., a first intron is between a first exon and a second exon. In some embodiments, a target portion is within an exon. In some embodiments, a target portion is within an intron. In some embodiments, a target portion comprises a portion of an exon and a portion of an intron.

[0153] In some embodiments, a target portion comprises a portion of a target nucleic acid (e.g., transcript) that encodes a polypeptide, e.g., a protein. In some embodiments, a target portion comprises a portion of an untranslated region (UTR), e.g., a 5’ UTR or a 3’ UTR. In some embodiments, a target portion comprises a portion of a 5 ’ UTR. In some embodiments, a target portion comprises a portion of a 3 ’ UTR.

[0154] In some embodiments, a target portion is or comprises a characteristic portion of a nucleic acid sequence (e.g., of a gene or a transcript thereof) which characteristic portion defines the nucleic acid sequence over others in a relevant organism; for example, the characteristic portion is not in other genomic nucleic acid sequences (e.g., genes or transcripts thereof) in a relevant organism (e.g., for a human gene or a transcript thereof, its characteristic portion is not in other human nucleic acid sequences or transcripts thereof). In some embodiments, a characteristic portion of a transcript defines that transcript over other transcripts in a relevant organism; for example, in some embodiments, the characteristic portion is not in transcripts that are transcribed from a different nucleic acid sequence (e.g., a different gene). In some embodiments, transcript variants from a nucleic acid sequence (e.g., mRNA variants of a gene) may share a common characteristic portion that defines them from transcripts of other nucleic acids, e.g., transcripts of other genes. In some embodiments, a characteristic portion in a transcript defines the transcript from other transcript(s) of the same nucleic acid sequence (e.g., a gene) and / or other alleles of the nucleic acid sequence. In some embodiments, a characteristic portion defines a particular allele (and / or transcripts thereof) over other allele(s) (and / or transcripts thereof). In some embodiments, a characteristic portion comprises sequences that are separated in a nucleic acid. In some embodiments, a characteristic portion is a contiguous stretch of nucleobases in a nucleic acid (a “characteristic sequence”). A characteristic portion or sequence may have various numbers of nucleobases. In some embodiments, there are about 10. 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more nucleobases in a characteristic portion or sequence; in some embodiments, there are about 10; in some embodiments, there are about 11 ; in some embodiments, there are about 12; in some embodiments, there are about 13; in some embodiments, there are about 14; in some embodiments, there are about 15; in some embodiments, there are about 16; in some embodiments, there are about 17; in some embodiments, there are about 18; in some embodiments, there are about 19; in some embodiments, there are about 20; in some embodiments, there are about 21; in some embodiments, there are about 22; in some embodiments, there are about 23; in some embodiments, there are about 24; in some embodiments, there are about 25; in some embodiments, there are about 25 or more.Attorney Docket No.: 2010794-3211

[0155] In some embodiments, an oligonucleotide comprises a sequence that is identical or complementary to a characteristic portion of a target nucleic acid. In some embodiments, an oligonucleotide comprises a sequence that is identical or complementary to a characteristic portion of a target transcript. In some embodiments, a base sequence of an antisense strand is identical or complementary to a characteristic portion of a target nucleic acid. In some embodiments, a base sequence of a sense strand is identical or complementary to a characteristic portion of a target nucleic acid. In some embodiments, a base sequence of an antisense strand is identical or complementary to a characteristic portion of a target transcript. In some embodiments, a base sequence of a sense strand is identical or complementary to a characteristic portion of a target transcript. In some embodiments, a base sequence of a sense strand is identical to a characteristic portion of a target transcript. In some embodiments, a base sequence of an antisense strand is complementary to a characteristic portion of a target transcript. In some embodiments, a characteristic portion is a characteristic sequence.

[0156] In some embodiments, an oligonucleotide comprises a sequence that is not identical or less than 100% complementary to a characteristic portion of a target nucleic acid. In some embodiments, an oligonucleotide comprises a sequence that is not identical or less than 100% complementary to a characteristic portion of a target transcript. In some embodiments, an oligonucleotide comprises a sequence that comprises one or more (e.g., 1, 2, 3 or more) mismatches as compared to a characteristic portion of a target nucleic acid. In some embodiments, an oligonucleotide comprises a sequence that comprises one or more (e.g., 1, 2, 3 or more) mismatches as compared to a characteristic portion of a target transcript. In some embodiments, a mismatch is at a 5’ end of an oligonucleotide. In some embodiments, a mismatch is at a 3’ end of an oligonucleotide. In some embodiments, a base sequence of an antisense strand is not identical or less than 100% complementary to a characteristic portion of a target nucleic acid. In some embodiments, a base sequence of an antisense strand comprises a sequence that is not identical or less than 100% complementary to a characteristic portion of a target transcript. In some embodiments, a base sequence of an antisense strand comprises a sequence that comprises one or more (e.g., 1, 2, 3 or more) mismatches as compared to a characteristic portion of a target nucleic acid. In some embodiments, a base sequence of an antisense strand comprises a sequence that comprises one or more (e.g., 1, 2, 3 or more) mismatches as compared to a characteristic portion of a target transcript. In some embodiments, a mismatch is at a 5’ end of an antisense strand. In some embodiments, a mismatch is at a 3’ end of an antisense strand. In some embodiments, a base sequence of a sense strand is not identical or less than 100% complementary to a characteristic portion of a target nucleic acid. In some embodiments, a base sequence of a sense strand comprises a sequence that is not identical or less than 100% complementary to a characteristic portion of a target transcript. In some embodiments, a base sequence of a sense strand comprises a sequence that comprises one or more (e.g., 1, 2, 3 or more) mismatches as compared to a characteristic portion of a targetAttorney Docket No.: 2010794-3211nucleic acid. In some embodiments, a base sequence of a sense strand comprises a sequence that comprises one or more (e.g., 1, 2, 3 or more) mismatches as compared to a characteristic portion of a target transcript. In some embodiments, a mismatch is at a 5’ end of a sense strand. In some embodiments, a mismatch is at a 3’ end of a sense strand.Lengths

[0157] As appreciated by those skilled in the art, oligonucleotides, and antisense strands and sense strands thereof, can be of various lengths to provide desired properties and / or activities for various uses. Many technologies for assessing, selecting and / or optimizing oligonucleotide length are available in the art and can be utilized in accordance with the present disclosure. As demonstrated herein, in many embodiments, provided oligonucleotides are of suitable lengths to provide desired reduction of their targets, e.g., transcripts, e.g., mRNA. As also demonstrated herein, in many embodiments, provided antisense strands are of suitable lengths to hybridize with their targets and reduce levels of their targets and / or a product thereof. In some embodiments, an antisense strand is sufficiently long enough to hybridize to a sense strand and vice-versa. In some embodiments, an oligonucleotide is long enough to recognize a target nucleic acid, e.g., a target transcript. In some embodiments, an oligonucleotide is sufficiently long to distinguish between a target nucleic acid and other nucleic acids (e.g., a nucleic acid having a base sequence which is not a target nucleic acid sequence) to reduce off-target effects. In some embodiments, an oligonucleotide is sufficiently short to reduce complexity of manufacture or production and to reduce cost of products.

[0158] In some embodiments, a base sequence of an oligonucleotide is about 10-100 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 10-50 nucleobases in length. In some embodiments, a bases sequence of an oligonucleotide is about 15-50 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 15-45 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 15-40 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 15-35 nucleobases in length.some embodiments, a base sequence of an oligonucleotide about 15-30 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 15-25 nucleobases in length.some embodiments, a base sequence of an oligonucleotide about 17-25 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 19-25 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 21-25 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 , 28, 29, or 30 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 15 nucleobases in length. In some embodiments, a base sequence of anAttorney Docket No.: 2010794-3211oligonucleotide is about 16 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 17 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 18 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 19 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 20 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 21 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 22 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 23 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 24 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 25 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 26 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 27 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 28 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 29 nucleobases in length. In some embodiments, a base sequence of an oligonucleotide is about 30 nucleobases in length.

[0159] In some embodiments, a base sequence of an antisense strand is about 10-50 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 15-50 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 15-45 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 15-40 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 15-35 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 15-30 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 15-25 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 17-25 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 19-25 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 21-25 nucleobases in length. In some embodiments, a base sequence of an antisense strand is 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 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 15 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 15 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 16 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 16 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 17 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 17Attorney Docket No.: 2010794-3211nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 18 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 18 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 19 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 19 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 20 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 20 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 21 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 21 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 22 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 22 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 23 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 23 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 24 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 24 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 25 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 25 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 26 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 26 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 27 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 27 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 28 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 28 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 29 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 29 nucleobases in length. In some embodiments, a base sequence of an antisense strand is at least about 30 nucleobases in length. In some embodiments, a base sequence of an antisense strand is about 30 nucleobases in length.

[0160] In some embodiments, a base sequence of a sense strand is about 10-50 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 15-50 nucleobases in length. In some embodiments, a base sequence of a sense strand i about 15-45 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 15-40 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 15-35 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 15-30 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 15-25 nucleobases in length. In someAttorney Docket No.: 2010794-3211embodiments, a base sequence of a sense strand is about 17-25 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 17-23 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 19-23 nucleobases in length. In some embodiments, abase sequence of a sense strand is 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 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 15 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 15 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 16 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 16 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 17 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 17 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 18 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 18 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 19 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 19 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 20 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 20 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 21 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 21 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 22 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 22 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 23 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 23 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 24 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 24 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 25 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 25 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 26 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 26 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 27 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 27 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 28 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 28 nucleobases in length. In some embodiments, a base sequence of a sense strand is at least about 29 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 29 nucleobases in length. In some embodiments,Attorney Docket No.: 2010794-3211a base sequence of a sense strand is at least about 30 nucleobases in length. In some embodiments, a base sequence of a sense strand is about 30 nucleobases in length.

[0161] In some embodiments, an oligonucleotide comprises an overhang on an antisense strand. In some embodiments, an oligonucleotide comprises an overhang on a sense strand. In some embodiments, an oligonucleotide comprises an overhang on an antisense strand and an overhang on a sense strand. In some embodiments, an overhang is about 1-5 nucleobases in length. In some embodiments, an overhang is about 1-4 nucleobases in length. In some embodiments, an overhang is about 1-3 nucleobases in length. In some embodiments, an overhang is about 2-5 nucleobases in length. In some embodiments, an overhang is about 2-4 nucleobases in length. In some embodiments, an overhang is about 1, 2, 3, 4, or 5 nucleobases in length. In some embodiments, an overhang is 1 nucleobase in length. In some embodiments, an overhang is 2 nucleobases in length. In some embodiments, an overhang is 3 nucleobases in length. In some embodiments, an overhang is 4 nucleobases in length. In some embodiments, an overhang is 5 nucleobases in length.Nucleobases

[0162] Various nucleobases may be utilized in provided oligonucleotides in accordance with the present disclosure. In some embodiments, a nucleobase is a natural nucleobase, the most commonly occurring ones being A, T, C, G and U. In some embodiments, a nucleobase is a modified nucleobase in that it is not A, T, C, G or U. In some embodiments, a nucleobase is optionally substituted A, T, C, G or U, or a substituted tautomer of A T, C, G or U. In some embodiments, a nucleobase is optionally substituted A, T, C, G or U, e.g., 5mC, 5 -hydroxymethyl C, etc. In some embodiments, a nucleobase is alkylsubstituted A, T, C, G or U. In some embodiments, a nucleobase is A. In some embodiments, a nucleobase is T. In some embodiments, a nucleobase is C. In some embodiments, a nucleobase is G. In some embodiments, a nucleobase is U. In some embodiments, a nucleobase is 5mC. In some embodiments, a nucleobase is substituted A, T, C, G or U. In some embodiments, a nucleobase is a substituted tautomer of A, T, C, G or U. In some embodiments, substitution protects certain functional groups in nucleobases to minimize undesired reactions during oligonucleotide synthesis. Suitable technologies for nucleobase protection in oligonucleotide synthesis are widely known in the art and may be utilized in accordance with the present disclosure. In some embodiments, modified nucleobases improves properties and / or activities of oligonucleotides. For example, in many cases, 5mC may be utilized in place of C to modulate certain undesired biological effects, e.g., immune responses. In some embodiments, when determining sequence identity, a substituted nucleobase having the same hydrogen-bonding pattern is treated as the same as the unsubstituted nucleobase, e.g., 5mC may be treated the same as C (e.g., an oligonucleotide having 5mC in place of C (e.g., AT5mCG) is considered to have the same base sequence as an oligonucleotide having CAttorney Docket No.: 2010794-3211at the corresponding location(s) (e.g., ATCG)).

[0163] In some embodiments, an oligonucleotide comprises one or more A, T, C, G or U. In some embodiments, an oligonucleotide comprises one or more optionally substituted A, T, C, G or U. In some embodiments, an oligonucleotide comprises one or more 5 -methylcytosine (5mC), 5-hydroxymethylcytosine, 5 -formylcytosine, or 5-carboxylcytosine. In some embodiments, an oligonucleotide comprises one or more 5mC. In some embodiments, each nucleobase in an oligonucleotide is independently selected from optionally substituted A, T, C, G and U, and optionally substituted tautomers of A, T, C, G and U. In some embodiments, each nucleobase in an oligonucleotide is independently optionally protected A, T, C, 5mC, G or U. In some embodiments, each nucleobase in an oligonucleotide is independently optionally substituted A, T, C, G or U. In some embodiments, each nucleobase in an oligonucleotide is selected from the group consisting of A, T, C, G, U, and 5mC.

[0164] In some embodiments, a nucleobase is isocytosine. In some embodiments, an oligonucleotide comprises one or more isocytosine. In some embodiments, an oligonucleotide comprises one or more optionally substituted isocytosine. In some embodiments, an oligonucleotide comprises one or more optionally protected isocytosine. In some embodiments, each nucleobase in an oligonucleotide is independently selected from A, T, C, G, U, and isocytosine. In some embodiments, each nucleobase in an oligonucleotide is independently selected from A, T, C, 5mC, G, U, and isocytosine. In some embodiments, each nucleobase in an oligonucleotide is independently optionally protected A, T, C, G, U, or isocytosine. In some embodiments, each nucleobase in an oligonucleotide is independently optionally protected A, T, C, 5mC, G, U, and isocytosine. In some embodiments, each nucleobase in an oligonucleotide is independently optionally substituted A, T, C, G, U, or isocytosine. In some embodiments, each nucleobase in an oligonucleotide is independently optionally substituted A, T, C, 5mC, G, U, and isocytosine. In some embodiments, each nucleobase in an oligonucleotide is selected from the group consisting of A, T, C, G, U, and isocytosine. In some embodiments, each nucleobase in an oligonucleotide is selected from the group consisting of A, T, C, G, U, 5mC, and isocytosine.

[0165] In some embodiments, a nucleobase is isoguanine. In some embodiments, an oligonucleotide comprises one or more isoguanine. In some embodiments, an oligonucleotide comprises one or more optionally substituted isoguanine. In some embodiments, an oligonucleotide comprises one or more optionally protected isoguanine. In some embodiments, each nucleobase in an oligonucleotide is independently selected from A, T, C, G, U, and isoguanine. In some embodiments, each nucleobase in an oligonucleotide is independently selected from A, T, C, 5mC, G, U, and isoguanine. In some embodiments, each nucleobase in an oligonucleotide is independently optionally protected A, T, C, G, U, or isoguanine. In some embodiments, each nucleobase in an oligonucleotide is independently optionally protected A, T, C, 5mC, G, U, and isoguanine. In some embodiments, each nucleobase in an oligonucleotide isAttorney Docket No.: 2010794-3211independently optionally substituted A, T, C, G, U, or isoguanine. In some embodiments, each nucleobase in an oligonucleotide is independently optionally substituted A, T, C, 5mC, G, U, and isocytosine. In some embodiments, each nucleobase in an oligonucleotide is selected from the group consisting of A, T, C, G, U, and isoguanine. In some embodiments, each nucleobase in an oligonucleotide is selected from the group consisting of A, T, C, G, U, 5mC, and isoguanine.

[0166] In some embodiments, each nucleobase in an oligonucleotide is independently selected from A, T, C, G, U, isocytosine, and isoguanine. In some embodiments, each nucleobase in an oligonucleotide is independently selected from A, T, C, 5mC, G, U, isocytosine, and isoguanine. In some embodiments, each nucleobase in an oligonucleotide is independently optionally protected A, T, C, G, U, isocytosine, or isoguanine. In some embodiments, each nucleobase in an oligonucleotide is independendy optionally protected A, T, C, 5mC, G, U, isocytosine, and isoguanine. In some embodiments, each nucleobase in an oligonucleotide is independently optionally substituted A, T, C, G, U, isocytosine, or isoguanine. In some embodiments, each nucleobase in an oligonucleotide is independently optionally substituted A, T, C, 5mC, G, U, isocytosine, and isocytosine. In some embodiments, each nucleobase in an oligonucleotide is selected from the group consisting of A, T, C, G, U, isocytosine, and isoguanine. In some embodiments, each nucleobase in an oligonucleotide is selected from the group consisting of A, T, C, G, U, 5mC, isocytosine, and isoguanine.

[0167] In some embodiments, a GNA nucleoside comprises an isocytosine nucleobase or an isoguanine nucleobase. In some embodiments, a GNA nucleoside comprises an isocytosine nucleobase. In some embodiments, a GNA nucleoside comprises an isoguanine nucleobase. As described herein, a GNA nucleoside comprising an isocytosine nucleobase or an isoguanine nucleobase is referred to as an GNA-iso nucleoside.

[0168] In some embodiments, a nucleobase is optionally substituted 2-aminopurine (2AP) or 2,6-diaminopurine (DAP). In some embodiments, a nucleobase is optionally substituted 2AP. In some embodiments, a nucleobase is optionally substituted DAP. In some embodiments, a nucleobase is 2AP. In some embodiments, a nucleobase is DAP.

[0169] In some embodiments, a nucleobase is a natural nucleobase or a modified nucleobase derived from a natural nucleobase. Examples include uracil, thymine, adenine, cytosine, and guanine optionally having their respective amino groups protected by acyl protecting groups, 2-fluorouracil, 2-fluorocytosine, 2-thiouracil, 5 -bromouracil, 5-iodouracil, 5 -fluorouracil, 2,6-diaminopurine, N6-methyladenine, 6-phenylpyrrolocytosine, 5-methylcytidine, azacytosine, N-ethylpiperidine 7’-EAA triazole modified adenine, N-ethylpiperidine 6’ -triazole modified adenine, pyrimidine analogs such as pseudoisocytosine and pseudouracil and other modified nucleobases such as 8-substituted purines, xanthine, or hypoxanthine (the latter two being the natural degradation products). Certain examples of modified nucleobases are disclosedAttorney Docket No.: 2010794-3211in Chiu and Rana. RNA. 2003 ;9(9): 1034- 1048, Limbach et al. Nucleic Acids Research. 1994;22( 12):2183-2196, Revankar and Rao, Comprehensive Natural Products Chemistry, vol. 7, 313; and Hu et al. Signal Transduct Target Ther. 2020;5(l):101.

[0170] In some embodiments, a modified nucleobase is a modified nucleobase known in the art. In some embodiments, modified nucleobases are expanded-size nucleobases in which one or more aryl and / or heteroaryl rings, such as phenyl rings, have been added.

[0171] Nucleobases may be protected during oligonucleotide synthesis. Various protection technologies are available and can be utilized in accordance with the present disclosure.

[0172] In some embodiments, a modified nucleobase is 5-substituted pyrimidines, 6- azapyrimidines, alkyl or alkynyl substituted pyrimidines, alkyl substituted purines, or N-2, N-6 and O-6 substituted purines. In certain embodiments, a modified nucleobase is selected form 2-aminopropyladenine, 5 -hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-N-methylguanine, 6-N- methyladenine, 2-propyladenine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl (-C=C-CHs) uracil, 5-propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothymine, 5 -ribosyluracil (pseudouracil), 4- thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, 8-aza and other 8-substituted purines, 5-halo, particularly 5-bromo, 5 -trifluoromethyl, 5-halouracil, and 5-halocytosine, 7-methylguanine, 7-methyladenine, 2-F-adenine, 2-aminoadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine, 3-deazaadenine, 6-N-benzoyladenine, 2-N-isobutyrylguanine, 4-N-benzoylcytosine, 4-N-benzoyluracil, 5-methyl 4-N-benzoylcytosine, 5-methyl 4-N-benzoyluracil, universal bases, hydrophobic bases, promiscuous bases, size-expanded bases, and fluorinated bases. In some embodiments, a modified nucleobase is a tricyclic pyrimidine, such as l,3-diazaphenoxazine-2-one, l,3-diazaphenothiazine-2-one or 9-(2-aminoethoxy)-l,3-diazaphenoxazine-2-one (G-clamp). In some embodiments, a modified nucleobase is one in which a purine or pyrimidine base is replaced with other heterocycles, for example, 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine or 2-pyrridone.

[0173] In some embodiments, a modified nucleobase is substituted. In some embodiments, a modified nucleobase is substituted such that it contains, e.g., heteroatoms, alkyl groups, or linking moieties connected to fluorescent moieties, biotin or avidin moieties, or other protein or peptides. In some embodiments, a modified nucleobase is a “universal base” that is not a nucleobase in the most classical sense, but that functions similarly to a nucleobase. One example of a universal base is 3-nitropyrrole.

[0174] In some embodiments, nucleosides that can be utilized in provided technologies comprise modified nucleobases and / or modified sugars, e.g., 4-acetylcytidine, 5-(carboxyhydroxylmethyl)uridine, 2 ’ -O-methylcytidine, 5 -carboxymethylaminomethyl-2-thiouridine, 5 -carboxymethylaminomethyluridine, dihydrouridine, 2’-O-methylpseudouridine, beta,D-galactosylqueosine, 2’-O-methylguanosine, N6-isopentenyladenosine, 1 -methyladenosine, 1-methylpseudouridine, 1 -methylguanosine, 1-methylinosine,Attorney Docket No.: 2010794-32112,2-dimethylguanosine, 2-methyladenosine, 2-methylguanosine, N7-methylguanosine, 3-methyl-cytidine, 5 -methylcytidine, 5 -hydroxymethylcytidine, 5 -formylcytosine, 5 -carboxyl cytosine, N6-methyladenosine, 7-methylguanosine, 5 -methylaminoethyluridine, 5-methoxyaminomethyl-2-thiouridine, beta,D-mannosylqueosine, 5 -methoxy carbonylmethyluridine, 5-methoxyuridine, 2-methylthio-N6-isopentenyladenosine, N-((9-beta,D-ribofuranosyl-2-methylthiopurine-6-yl)carbamoyl)threonine, N-((9-beta,D-ribofuranosylpurine-6-yl)-N-methylcarbamoyl)threonine, uridine-5-oxyacetic acid methylester, uridine-5-oxyacetic acid (v), pseudouridine, queosine, 2-thiocytidine, 5-methyl-2-thiouridine, 2-thiouridine, 4-thiouridine, 5-methyluridine, 2’ -O-methyl-5 -methyluridine, and 2’-O-methyluridine.

[0175] In some embodiments, a nucleobase, e.g., a modified nucleobase comprises one or more biomolecule binding moieties such as e.g., antibodies, antibody fragments, biotin, avidin, streptavidin, receptor ligands, or chelating moieties. In other embodiments, a nucleobase is 5 -bromouracil, 5-iodouracil, or 2,6-diaminopurine. In some embodiments, a nucleobase comprises substitution with a fluorescent or biomolecule binding moiety. In some embodiments, a substituent is a fluorescent moiety. In some embodiments, a substituent is biotin or avidin.

[0176] In some embodiments, a nucleobase is one described in US 9394333, US 9744183, US 9605019, US 9598458, US 9982257, US 10160969, US 10479995, US 2020 / 0056173, US 2018 / 0216107, US 2019 / 0127733, US 10450568, US 2019 / 0077817, US 2019 / 0249173, US 2019 / 0375774, US 2012 / 0225034, WO 2018 / 223056, WO 2018 / 223073, WO 2018 / 223081, WO 2018 / 237194, WO 2019 / 032607, WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784, WO 2019 / 032612, WO 2013 / 016352, WO 2016 / 115490, WO 2020 / 028134, WO 2022 / 020106, WO 2022 / 051332, WO 2022 / 056266, WO 2022 / 147207, WO 2022 / 159712, WO 2023 / 018705, WO 2023 / 168202, WO 2023 / 283629, WO 2023 / 288289, and / or WO 2024 / 011135, the nucleobases of each of which is incorporated herein by reference.Sugars

[0177] Various sugars, including modified sugars, can be utilized in accordance with the present disclosure. In some embodiments, the present disclosure provides sugar modifications and patterns thereof optionally in combination with other structural elements (e.g., nucleobase modifications and patterns thereof, internucleotidic linkage modifications and patterns thereof, etc.) that when incorporated into oligonucleotides can provide improved properties and / or activities.

[0178] The most common naturally occurring nucleosides comprise ribose sugars (e.g., in RNA) or deoxyribose sugars (e.g., in DNA) linked to the nucleobases adenosine (A), cytosine (C), guanine (G), thymine (T) or uracil (U). In some embodiments, a sugar is a natural DNA sugar (in DNA nucleic acids orAttorney Docket No.: 2010794-3211oligonucleotides, having the structure of, wherein a nucleobase is attached to the 1 ’ position, and the 3’ and 5’ positions are connected to internucleotidic linkages (as appreciated by those skilled in the art, if at the 5’-end of an oligonucleotide, the 5’ position may be connected to a 5’-end group (e.g., -OH), and if at the 3’-end of an oligonucleotide, the 3’ position may be connected to a 3’-end group (e.g., -OH). In some embodiments, a sugar is a natural RNA sugar (in RNA nucleic acids or oligonucleotides, havingthe structure of OH;wherein a nucleobase is attached to the 1’ position, and the 3’ and 5’ positions are connected to internucleotidic linkages (as appreciated by those skilled in the art, if at the 5 ’ -end of an oligonucleotide, the 5’ position may be connected to a 5’-end group (e.g., -OH), and if at the 3’-end of an oligonucleotide, the 3’ position may be connected to a 3 ’-end group (e.g., -OH). In some embodiments, a sugar is a modified sugar in that it is not a natural DNA sugar or a natural RNA sugar. Among other things, modified sugars may provide improved stability and / or affinity. In some embodiments, modified sugars can be utilized to alter and / or optimize one or more hybridization characteristics. In some embodiments, modified sugars can be utilized to alter and / or optimize target recognition. In some embodiments, modified sugars can be utilized to optimize Tin. In some embodiments, modified sugars can be utilized to improve oligonucleotide activities.

[0179] Sugars can be bonded to internucleotidic linkages at various positions. As non-limiting examples, internucleotidic linkages can be bonded to the 2’, 3’, 4’ or 5’ positions of sugars. As most commonly in natural nucleic acids, an internucleotidic linkage typically connects with one sugar at the 5’ position and another sugar at the 3’ position unless otherwise indicated.

[0180] In some embodiments, a sugar is an optionally substituted natural DNA or RNA sugar. In some 514'1embodiments, a sugar is optionally substitutedIn some embodiments, the 2’ position is 514'1 0optionally substituted. In some embodiments, a sugar is. In some embodiments, a sugar has R55S^O- 5' 02-~R2Scu eR2Sthe stru t r of ,2s wherein each of Rls, R2s, R3s, R4s, and R5sisAttorney Docket No.: 2010794-3211independently -H, a suitable substituent or suitable sugar modification (e.g., those described in US 9394333, US 9744183, US 9605019, US 9982257, US 20170037399, US 20180216108, US 20180216107, US 9598458, WO 2017 / 062862, WO 2018 / 067973, WO 2017 / 160741, WO 2017 / 192679, WO 2017 / 210647, WO 2018 / 098264, WO 2018 / 022473, WO 2018 / 223056, WO 2018 / 223073, WO 2018 / 223081, WO 2018 / 237194, WO 2019 / 032607, WO 2019 / 032612, WO 2019 / 055951, and / or WO 2019 / 075357, the substituents, sugar modifications, descriptions of Rls, R2s, R3s, R4s, and R5s, and modified sugars of each of which are independently incorporated herein by reference). In some embodiments, each of Rls, R2S, R3S, R4S, and R5sis independently -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -Ls-R’, -LS-OR’, -LS-SR’, -LS-N(R’)2, - O— Ls- OR’, -O-LS-SR’, or -O-LS-N(R’)2, wherein each R’ is independently -H or an optionally substituted group selected from Ci-io aliphatic, Ce-u aryl, Ci-io heteroaliphatic having 1-5 heteroatoms, 5-10 membered heteroaryl having 1-5 heteroatoms and 3-10 membered heterocyclyl having 1-4 heteroatoms, or two or more R’ groups are taken together with their intervening atoms to from an optionally substituted 3-10 membered ring having 0-5 heteroatoms in addition to the intervening atoms. Independently as described herein, and Lsis a covalent bond or optionally substituted bivalent Ci-e aliphatic5' 4' R I \4!-sor heteroaliphatic having 1-4 heteroatoms. In some embodiments, a sugar has thestructure ofRIn some embodiments, R4sis -H. In some embodiments, a sugar has the structure of wherein R2sis -H, halogen, or -OR, wherein R is optionally substituted Ci-6 aliphatic. In some embodiments, R2sis -H. In some embodiments, R2sis -F. In some embodiments, R2sis -Ome. In some embodiments, R2sis -OCH2CH2Ome.CK2L

[0181] In some embodiments, a sugar has the structureof >2s , wherein R2sand R4sare taken together to form -Ls-, wherein Lsis a covalent bond or optionally substituted bivalent Ci-6 aliphatic or heteroaliphatic having 1-4 heteroatoms. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen or sulfur). In some embodiments, Lsis optionally substituted C2-O-CH2-C4. In some embodiments, Lsis C2-O-CH2-C4. In some embodiments, Lsis C2-O-I-CH(CH2CH3)-C4. In some embodiments, Lsis C2-O-(S)-CH(CH2CH3)-C4.Attorney Docket No.: 2010794-3211

[0182] In some embodiments, a sugar has the structureof wherein R2sis -H, halogen, or -OR, wherein R is optionally substituted Ci-6 aliphatic. In some embodiments, R2sis -H. In some embodiments, R2sis -F.

[0183] In some embodiments, a modified sugar contains one or more substituents at the 2’ position ( (typically one substituent, and often at the axial position or R2s) independently selected from -F; -CF?, -CN, -N3, -NO, -NO2, -OR’, -SR’, or -N(R’)2, wherein each R’ is independently described in the present disclosure, and in some embodiments, optionally substituted Cuo aliphatic; -0-(Ci-Cio alkyl), -S-(Ci-C10 alkyl), -NH-(Ci-Cio alkyl), or -N(Ci-Cw alkyl)2; -0-(C2-Cio alkenyl), -S-(C2-Cio alkenyl), -NH-(C2-Cio alkenyl), or -N(C2-Cio alkenyl)2; -0-(C2-Cio alkynyl), -S-(C2-Cio alkynyl), -NH-(C2-CIO alkynyl), or -N(C2-CW alkynyl)2; or -O — (C1-C10 alkylene)-0 — (C1-C10 alkyl), -O-(Ci-Cw alkylene)-NH-(Ci-C alkyl) or -O-(Ci-C alkylene)-NH(C1-C10alkyl)2, -NH-(Ci-Cw alkylene)-O-(Ci-C alkyl), or -N(Ci-Cio alkyl)-(Ci-Cio alkylene)-0-(Ci-Cio alkyl), wherein each of the alkyl, alkylene, alkenyl and alkynyl is independently and optionally substituted. In some embodiments, a substituent is - O(CH2)nOCH3, -O(CH2)nNH2, MOE, DMAOE, or DMAEOE, wherein n is from 1 to about 10.

[0184] In some embodiments, a modified sugar is a natural RNA sugar whose 2’ -OH is replaced with a group selected from -F, -CF3, -CN, -N3, -NO, -NO2, -OR’, -SR’, or -N(R’)2, wherein each R’ is independently described in the present disclosure; -0-(Ci-Cio alkyl), -S-(Ci-Cio alkyl), -NH-(Ci-Cio alkyl), or -N(Ci-Cio alkyl)2; -0-(C2-Cio alkenyl), -S-(C2-Cio alkenyl), -NH-(C2-C10alkenyl), or -N(C2-C10 alkenyl)2; -0-(C2-Cio alkynyl), -S-(C2-Cio alkynyl), -NH-(C2-CIO alkynyl), or -N(C2-CIO alkynyl)2; or -O — (C1-C10 alkylene)-0 — (C1-C10 alkyl), -O-(Ci-Cw alkylene)-NH-(Ci-Cw alkyl) or -0-(Ci-Cio alkylene)-NH(Ci-Cio alkyl)2, -NH-(Ci-Cw alkylene)-0-(Ci-Cio alkyl), or -N(Ci-Cio alkyl)-(Ci-Cio alkylene)-0-(Ci-Cio alkyl), wherein each of the alkyl, alkylene, alkenyl and alkynyl is independently and optionally substituted. In some embodiments, the 2’-OH is replaced with -H (deoxyribose). In some embodiments, the 2’-OH is replaced with -F. In some embodiments, the 2’-OH is replaced with -OR’ . In some embodiments, the 2’-OH is replaced with -Ome. In some embodiments, the 2’-OH is replaced with -OCH2CH2Ome.

[0185] In some embodiments, a sugar modification is a 2’ -modification. In some embodiments, a 2’-modification is a 2’-ORsmodification. In some embodiments, Rsis optionally substituted C1.4 aliphatic. In some embodiments, Rsis optionally substituted Ci.g alkyl. In some embodiments, a modification is 2’-OMe. In some embodiments, a modification is 2’-MOE. In some embodiments, a 2 ’-modification is -F. In some embodiments, a 2 ’-modification is S-cEt. In some embodiments, a 2 ’-modification is FANA.Attorney Docket No.: 2010794-3211

[0186] In some embodiments, a modified sugar is an LNA sugar. In some embodiments, a modified 5'sugar has a structureof '"i1"

[0187] In some embodiments, a sugar modification replaces a sugar moiety with another cyclic or acyclic moiety. Examples of such moieties are widely known in the art, e.g., those in morpholino, glycol nucleic acid (GNA), peptide nucleic acid (PNA), threose nucleic acid (TNA), unlocked nucleic acid (UNA), etc., and may be utilized in accordance with the present disclosure. In some embodiments, a modified sugaris a morpholino sugar. In some embodiments, a modified sugar has a structureof . In some embodiments, a modified sugar is a GNA sugar. In some embodiments, a modified sugar has a structureof i . In some embodiments, a modified sugar is a TNA sugar. In some embodiments, a modifiedsugar has a structureof . In some embodiments, a modified sugar is a UNA sugar. In someembodiments, a modified sugar has a structureof . wherein R2sis -H, -OH, halogen, or -OR, wherein R is optionally substituted Ci-6 aliphatic. In some embodiments, R2sis -OH. In some embodiments, R2sis -H. In some embodiments, R2sis -F. In some embodiments, R2sis -OMe. In some embodiments, R2sis -OCH2CH2OMe.

[0188] In some embodiments, one or more sugars of an oligonucleotide are independently modified. In some embodiments, each sugar of an oligonucleotide or a portion thereof is independently modified. In some embodiments, a modified sugar comprises a 2’ -modification. In some embodiments, each modified sugar independently comprises a 2’ -modification. In some embodiments, a 2 ’-modification is 2’-ORs, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a 2’ -modification is a 2’ -OMe modification. In some embodiments, a 2 ’-modification is a 2’-M0E modification. In some embodiments, a 2 ’-modification is 2’-F. In some embodiments, a modified sugar is an LNA sugar. In some embodiments, a modified sugar comprises a GNA sugar. In some embodiments, a modified sugar comprises a UNA sugar. In some embodiments, each sugar modification is independently a 2’ -modification. In some embodiments, each sugar modification is 2’-OMe, 2’-M0E, or 2’-F. In some embodiments, each sugar modification isAttorney Docket No.: 2010794-32112’-0Me or 2’-F.

[0189] As those skilled in the art will appreciate, modifications of sugars, nucleobases, internucleotidic linkages, etc. can and are often utilized in combination in oligonucleotides, e.g., see various oligonucleotides described herein, e.g., various oligonucleotides in Table 1 (e.g., Table 1A, Table IB, Table 1C), Table 3, and Table 4. Various additional sugars useful for preparing oligonucleotides or analogs thereof are known in the art and may be utilized in accordance with the present disclosure. See, e.g., Hu et al. Signal Transduct Target Ther. 2020;5( 1) : 101 , which is hereby incorporated by reference in its entirety.

[0190] In some embodiments, the present disclosure provides patterns of sugar modifications that can be used in, e.g., an oligonucleotide, an antisense strand, or a sense strand. Provided patterns of sugar modifications can be used to provide certain properties to an oligonucleotide, e.g., increased activities. Certain patterns of sugar modifications may also provide decreased toxicity (e.g., genotoxicity, mitochondrial toxicity).

[0191] In some embodiments, a pattern of sugar modifications comprises one or more modified sugars, e.g., 2’-F modified sugars, 2’-OMe modified sugars, 2’ -MOE modified sugars, GNA sugars, UNA sugars. In some embodiments, a pattern of sugar modifications comprises one or more natural sugars, e.g., DNA sugars. In some embodiments, a pattern of sugar modifications comprises one or more modified sugars, e.g., 2’-F modified sugars, 2’-OMe modified sugars, 2’-M0E modified sugars, GNA sugars, UNA sugars and one or more natural sugars, e.g., DNA sugars.

[0192] In some embodiments, a pattern of sugar modifications comprises one or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 2 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 3 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 4 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 5 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 6 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 7 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 8 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 9 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 10 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 11 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 12 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 13 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 14 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 15 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 16 or more 2’-F modified sugars. In someAttorney Docket No.: 2010794-3211embodiments, a pattern of sugar modifications comprises 17 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 18 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 19 or more 2’-F modified sugars. In some embodiments, a pattern of sugar modifications comprises 20 or more 2’-F modified sugars.

[0193] In some embodiments, a pattern of sugar modifications comprises one or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 2 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-e aliphatic. In some embodiments, a pattern of sugar modifications comprises 3 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 4 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 5 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 6 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-e aliphatic. In some embodiments, a pattern of sugar modifications comprises 7 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 8 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 9 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 10 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-e aliphatic. In some embodiments, a pattern of sugar modifications comprises 11 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 12 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 13 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 14 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 15 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 16 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 17 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 18 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 19 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6 aliphatic. In some embodiments, a pattern of sugar modifications comprises 20 or more 2’-ORsmodified sugars, wherein Rsis optionally substituted Ci-6Attorney Docket No.: 2010794-3211aliphatic.

[0194] In some embodiments, a pattern of sugar modifications comprises one or more 2’-OMe modified sugars. In some embodiments, a pattern of sugar modifications comprises 2 or more 2’-0Me modified sugars. In some embodiments, a pattern of sugar modifications comprises 3 or more 2’-0Me modified sugars. In some embodiments, a pattern of sugar modifications comprises 4 or more 2’-0Me modified sugars. In some embodiments, a pattern of sugar modifications comprises 5 or more 2’-0Me modified sugars. In some embodiments, a pattern of sugar modifications comprises 6 or more 2’-0Me modified sugars. In some embodiments, a pattern of sugar modifications comprises 7 or more 2’-0Me modified sugars. In some embodiments, a pattern of sugar modifications comprises 8 or more 2’-0Me modified sugars. In some embodiments, a pattern of sugar modifications comprises 9 or more 2’-0Me modified sugars. In some embodiments, a pattern of sugar modifications comprises 10 or more 2’-OMe modified sugars. In some embodiments, a pattern of sugar modifications comprises 11 or more 2’-OMe modified sugars. In some embodiments, a pattern of sugar modifications comprises 12 or more 2’-OMe modified sugars. In some embodiments, a pattern of sugar modifications comprises 13 or more 2’-OMe modified sugars. In some embodiments, a pattern of sugar modifications comprises 14 or more 2’-OMe modified sugars. In some embodiments, a pattern of sugar modifications comprises 15 or more 2’-OMe modified sugars. In some embodiments, a pattern of sugar modifications comprises 16 or more 2’-OMe modified sugars. In some embodiments, a pattern of sugar modifications comprises 17 or more 2’-OMe modified sugars. In some embodiments, a pattern of sugar modifications comprises 18 or more 2’-OMe modified sugars. In some embodiments, a pattern of sugar modifications comprises 19 or more 2’-OMe modified sugars. In some embodiments, a pattern of sugar modifications comprises 20 or more 2’-OMe modified sugars.

[0195] In some embodiments, a pattern of sugar modifications comprises one or more GNA sugars. In some embodiments, a pattern of sugar modifications comprises 2 or more GNA sugars. In some embodiments, a pattern of sugar modifications 3 or more GNA sugars. In some embodiments, a pattern of sugar modifications comprises 4 or more GNA sugars. In some embodiments, a pattern of sugar modifications comprises 5 or more GNA sugars.

[0196] In some embodiments, a pattern of sugar modifications comprises one or more UNA sugars. In some embodiments, a pattern of sugar modifications comprises 2 or more UNA sugars. In some embodiments, a pattern of sugar modifications comprises 3 or more UNA sugars. In some embodiments, a pattern of sugar modifications comprises 4 or more UNA sugars. In some embodiments, a pattern of sugar modifications comprises 5 or more UNA sugars.

[0197] In some embodiments, a pattern of sugar modifications comprises one or more natural DNA sugars, e.g., one or more sugars comprising 2’ -OH. In some embodiments, a pattern of sugar modificationsAttorney Docket No.: 2010794-3211comprises 2 or more natural DNA sugars. In some embodiments, a pattern of sugar modifications comprises 3 or more natural DNA sugars. In some embodiments, a pattern of sugar modifications comprises 4 or more natural DNA sugars. In some embodiments, a pattern of sugar modifications comprises 5 or more natural DNA sugars.

[0198] In some embodiments, a pattern of sugar modifications comprises mN fN (mN), fN mN (fN)b (mN)c fN mN fN (mN)d fN (mN)e, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, and each of a, b, c, d, and e are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN), fN mN (fN)b (mN), fN mN fN (mN)d fN (mN),, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, and each of a, b, c, d, and e are independently an integer of 1 or greater.

[0199] In some embodiments, an oligonucleotide comprises mN fN (mN), fN mN (fN)b (mN)cfN mN fN (mN)d fN (mN)e, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, and each of a, b, c, d, and e are independendy an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN), fN mN (fN)b (mN), fN mN fN (mN)d fN (mN),, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of a, b, c, d, and e are independently an integer of 1 or greater. In some embodiments, an antisense strand comprises mN fN (mN), fN mN (fN)b (mN), fN mN fN (mN)d fN (mN),, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, and each of a, b, c, d, and e are independently an integer of 1 or greater. In some embodiments, an antisense strand is mN fN (mN), fN mN (fN)b (mN), fN mN fN (mN)d fN (mN)c, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, and each of a, b, c, d, and e are independently an integer of 1 or greater.

[0200] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN), fN mN (fN)b (mN), fN mN fN (mN)a fN (mN)„ wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, and each of a, b, c, d, and e are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN), fN mN (fN)b (mN), fN mN fN (mN)d fN (mN),, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, and each of a, b, c, d, and e are independently an integer of 1 or greater.

[0201] In some embodiments, a is 1-10. In some embodiments, a is 2-10. In some embodiments, a is 1-5. In some embodiments, a is 2-5. In some embodiments, a is an integer selected from 2, 3, 4, 5, 6, or 7. In some embodiments, a is an integer selected from 2, 3, 4, or 5. In some embodiments, a is 1. In some embodiments, a is 2. In some embodiments, a is 3. In some embodiments, a is 4. In some embodiments, a is 5. In some embodiments, a is 6. In some embodiments, a is 7. In some embodiments, a is 8. In someAttorney Docket No.: 2010794-3211embodiments, a is 9. In some embodiments, a is 10.

[0202] In some embodiments, b is 1-10. In some embodiments, b is 2-10. In some embodiments, b is 1-5. In some embodiments, b is 2-5. In some embodiments, b is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, b is an integer selected from 2, 3, 4, or 5. In some embodiments, b is 1. In some embodiments, b is 2. In some embodiments, b is 3. In some embodiments, b is 4. In some embodiments, b is 5. In some embodiments, b is 6. In some embodiments, b is 7. In some embodiments, b is 8. In some embodiments, b is 9. In some embodiments, b is 10.

[0203] In some embodiments, c is 1-10. In some embodiments, c is 2-10. In some embodiments, c is 1-5. In some embodiments, c is 2-5. In some embodiments, c is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, c is an integer selected from 2, 3, 4, or 5. In some embodiments, c is 1. In some embodiments, c is 2. In some embodiments, c is 3. In some embodiments, c is 4. In some embodiments, c is 5. In some embodiments, c is 6. In some embodiments, c is 7. In some embodiments, c is 8. In some embodiments, c is 9. In some embodiments, c is 10.

[0204] In some embodiments, d is 1-10. In some embodiments, d is 2-10. In some embodiments, d is 1-5. In some embodiments, d is 2-5. In some embodiments, d is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, d is an integer selected from 2, 3, 4, or 5. In some embodiments, d is 1. In some embodiments, d is 2. In some embodiments, d is 3. In some embodiments, d is 4. In some embodiments, dis 5. In some embodiments, d is 6. In some embodiments, d is 7. In some embodiments, d is 8. In some embodiments, d is 9. In some embodiments, d is 10.

[0205] In some embodiments, e is 1-10. In some embodiments, e is 2-10. In some embodiments, e is 1-5. In some embodiments, e is 2-5. In some embodiments, e is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, e is an integer selected from 2, 3, 4, or 5. In some embodiments, e is 1. In some embodiments, e is 2. In some embodiments, e is 3. In some embodiments, e is 4. In some embodiments, e is 5. In some embodiments, e is 6. In some embodiments, e is 7. In some embodiments, e is 8. In some embodiments, e is 9. In some embodiments, e is 10.

[0206] In some embodiments, a sum of a, b, c, d, and e is not greater than 30. In some embodiments, a sum of a, b, c, d, and e is not greater than 25. In some embodiments, a sum of a, b, c, d, and e is not greater than 24. In some embodiments, a sum of a, b, c, d, and e is not greater than 23. In some embodiments, a sum of a, b, c, d, and e is not greater than 22. In some embodiments, a sum of a, b, c, d, and e is not greater than 21. In some embodiments, a sum of a, b, c, d, and e is not greater than 20. In some embodiments, a sum of a, b, c, d, and e is not greater than 19. In some embodiments, a sum of a, b, c, d, and e is not greater than 18. In some embodiments, a sum of a, b, c, d, and e is not greater than 17. In some embodiments, a sum of a, b, c, d, and e is not greater than 16. In some embodiments, a sum of a, b, c, d, and e is not greater than 15. In some embodiments, a sum of a, b, c, d, and e is not greater than 14. In someAttorney Docket No.: 2010794-3211embodiments, a sum of a, b, c, d, and e is not greater than 13. In some embodiments, a sum of a, b, c, d, and e is not greater than 12. In some embodiments, a sum of a, b, c, d, and e is not greater than 11. In some embodiments, a sum of a, b, c, d, and e is not greater than 10.

[0207] In some embodiments, each of a, b, c, d, and e is independently 1-10. In some embodiments, each of a, b, c, d, and e is independently 2-10. In some embodiments, each of a, b, c, d, and e is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of b, c, d, and e is independently an integer selected from 2, 3, 4, or 5. In some embodiments, each of b, c, d, and e is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, d, and e is independently an integer selected from 2, 3, 4, or 5. In some embodiments, each of b, d, and e is independently an integer selected from 2, 3. 4, 5, 6, 7, or 8. In some embodiments, each of b, d, and e is independently an integer selected from 2, 3, 4, or 5.

[0208] In some embodiments, a is 3, b is 2, c is 4, d is 3, and e is 3. In some embodiments, a is 1, b is 2, c is 4, d is 3, and e is 3.

[0209] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

[0210] In some embodiments, an oligonucleotide comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN f mN mN mN. In some embodiments, an oligonucleotide is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

[0211] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

[0212] In some embodiments, a pattern of sugar modifications comprises mN fN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

[0213] In some embodiments, an oligonucleotide comprises mN fN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an oligonucleotide is mN fN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. InAttorney Docket No.: 2010794-3211some embodiments, an antisense strand is mN fN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

[0214] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

[0215] In some embodiments, an oligonucleotide comprises mN * fN * mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * fN * mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0216] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, , wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an and sense strand that is mN * fN * mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0217] In some embodiments, a pattern of sugar modifications comprises fN mN fN (mN)gfN mN (fN)h (mN); fN (mN)j, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of g, h, i, and j are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is fN mN fN (mN)gIN mN (fN)h (mN)i fN (mN)j, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of g, h, i, and j are independently an integer of 1 or greater.

[0218] In some embodiments, an oligonucleotide comprises fN mN fN (mN)gfN mN (fN)h (mN)i fN (mN)j, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of g, h, i, and j are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is fN mN fN (mN)gfN mN (fN)h (mN)i fN (mN)j, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of g, h, i, and j are independently an integer of 1 or greater. In some embodiments, a sense strand comprises fN mN fN (mN)gfN mN (fN)h (mN)i fN (mN)j, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of g, h, i, and j are independently an integer of 1 or greater. InAttorney Docket No.: 2010794-3211some embodiments, a sense strand is fN mN fN (mN)gfN mN (fN)h (mN), fN (mN)j, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of g, h, i, and j are independently an integer of 1 or greater.

[0219] In some embodiments, an oligonucleotide comprises a sense strand comprising fN mN fN (mN)g fN mN (fN)h (mN)i fN (niN)j, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, and each of g, h, i, and j are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises a sense strand that is fN mN fN (mN)gfN mN (fN)h (mN)i fN (mN)j, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of g, h, i, and j are independently an integer of 1 or greater.

[0220] In some embodiments, g is 1-10. In some embodiments, g is 2-10. In some embodiments, g is 1-5. In some embodiments, g is 2-5. In some embodiments, g is an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, g is an integer selected from 2, 3, 4, 5, or 6. In some embodiments, g is 1. In some embodiments, g is 2. In some embodiments, g is 3. In some embodiments, g is 4. In some embodiments, g is 5. In some embodiments, g is 6. In some embodiments, g is 7. In some embodiments, g is 8. In some embodiments, g is 9. In some embodiments, g is 10.

[0221] In some embodiments, h is 1-10. In some embodiments, h is 2-10. In some embodiments, h is 1-5. In some embodiments, h is 2-5. In some embodiments, h is an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, h is an integer selected from 2, 3, 4, 5, or 6. In some embodiments, h is 1. In some embodiments, h is 2. In some embodiments, h is 3. In some embodiments, h is 4. In some embodiments, h is 5. In some embodiments, h is 6. In some embodiments, h is 7. In some embodiments, h is 8. In some embodiments, h is 9. In some embodiments, h is 10.

[0222] In some embodiments, i is 1-10. In some embodiments, i is 2-10. In some embodiments, i is 1-5. In some embodiments, i is 2-5. In some embodiments, i is an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, i is an integer selected from 2, 3, 4, 5, or 6. In some embodiments, i is 1. In some embodiments, i is 2. In some embodiments, i is 3. In some embodiments, i is 4. In some embodiments, i is 5. In some embodiments, i is 6. In some embodiments, i is 7. In some embodiments, i is 8. In some embodiments, i is 9. In some embodiments, i is 10.

[0223] In some embodiments, j is 1-10. In some embodiments, j is 2-10. In some embodiments, j is 1-5. In some embodiments, j is 2-5. In some embodiments, j is an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, j is an integer selected from 2, 3, 4, 5, or 6. In some embodiments, j is 1. In some embodiments, j is 2. In some embodiments, j is 3. In some embodiments, j is 4. In some embodiments, j is 5. In some embodiments, j is 6. In some embodiments, j is 7. In some embodiments, j is 8. In some embodiments,] is 9. In some embodiments,] is 10.

[0224] In some embodiments, a sum of g, h, i, and j is not greater than 30. In some embodiments, aAttorney Docket No.: 2010794-3211sum of g, h, i, and j is not greater than 25. In some embodiments, a sum of g, h, i, and j is not greater than 24. In some embodiments, a sum of g, h, i, and j is not greater than 23. In some embodiments, a sum of g, h, i, and j is not greater than 22. In some embodiments, a sum of g, h, i, and j is not greater than 21. In some embodiments, a sum of g, h, i, and j is not greater than 20. In some embodiments, a sum of g, h, i, and j is not greater than 19. In some embodiments, a sum of g, h, i, and j is not greater than 18. In some embodiments, a sum of g, h, i, and j is not greater than 17. In some embodiments, a sum of g, h, i, and j is not greater than 16. In some embodiments, a sum of g, h, i, and j is not greater than 15. In some embodiments, a sum of g, h, i, and j is not greater than 14. In some embodiments, a sum of g, h, i, and j is not greater than 13. In some embodiments, a sum of g, h, i, and j is not greater than 12. In some embodiments, a sum of g, h, i, and j is not greater than 11. In some embodiments, a sum of g, h, i, and j is not greater than 10.

[0225] In some embodiments, each of g, h, i, and j is independently 1-10. In some embodiments, each of g, h, i, and j is independently 2-10. In some embodiments, each of g, h, i, and j is independently an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, each of g, h, i, and j is independently an integer selected from 2, 3, 4, 5, or 6.

[0226] In some embodiments, g is 3, h is 3, i is 3. and j is 6.

[0227] In some embodiments, a pattern of sugar modifications comprises fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN. In some embodiments, a pattern of sugar modifications is fN mN fN mN mN mN fN mN fN fN f mN mN mN fN mN mN mN mN mN mN.

[0228] In some embodiments, an oligonucleotide comprises fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN. In some embodiments, an oligonucleotide is fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN. In some embodiments, a sense strand comprises fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN. In some embodiments, a sense strand is fN mN fN mN mN mN IN mN fN fN fN mN mN mN fN mN mN mN mN mN mN.

[0229] In some embodiments, an oligonucleotide comprises a sense strand comprising fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises a sense strand that is fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN.

[0230] In some embodiments, a pattern of sugar modifications comprises fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, a pattern of sugar modifications is fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.Attorney Docket No.: 2010794-3211

[0231] In some embodiments, an oligonucleotide comprises fN* mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, a sense strand comprises fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN. In some embodiments, a sense strand is fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0232] In some embodiments, an oligonucleotide comprises a sense strand comprising fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises a sense strand that is fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0233] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN)a fN mN (fN)b (mN)cfN mN fN (mN)d fN (mN)eand a sense strand comprising fN mN fN (mN)gfN mN (fN)h (mN)i fN (mN)j, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of a, b, c, d, e, g, h, i, and j are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN), fN mN (fN)t (mN)c fN mN fN (mN)d fN (mN)eand a sense strand that is fN mN fN (mN). fN mN (fN)h (mN), fN (mN)j, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of a, b, c, d, e, g, h, i, and j are independently an integer of 1 or greater.

[0234] In some embodiments, a is 1-10. In some embodiments, a is 2-10. In some embodiments, a is 1-5. In some embodiments, a is 2-5. In some embodiments, a is an integer selected from 2, 3, 4, 5, 6, or 7. In some embodiments, a is an integer selected from 2, 3, 4, or 5. In some embodiments, a is 1. In some embodiments, a is 2. In some embodiments, a is 3. In some embodiments, a is 4. In some embodiments, a is 5. In some embodiments, a is 6. In some embodiments, a is 7. In some embodiments, a is 8. In some embodiments, a is 9. In some embodiments, a is 10.

[0235] In some embodiments, b is 1-10. In some embodiments, b is 2-10. In some embodiments, b is 1-5. In some embodiments, b is 2-5. In some embodiments, b is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, b is an integer selected from 2, 3, 4, or 5. In some embodiments, b is 1. In some embodiments, b is 2. In some embodiments, b is 3. In some embodiments, b is 4. In some embodiments, b is 5. In some embodiments, b is 6. In some embodiments, b is 7. In some embodiments, b is 8. In some embodiments, b is 9. In some embodiments, b is 10.

[0236] In some embodiments, c is 1-10. In some embodiments, c is 2-10. In some embodiments, c isAttorney Docket No.: 2010794-32111-5. In some embodiments, c is 2-5. In some embodiments, c is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, c is an integer selected from 2, 3, 4, or 5. In some embodiments, c is 1. In some embodiments, c is 2. In some embodiments, c is 3. In some embodiments, c is 4. In some embodiments, c is 5. In some embodiments, c is 6. In some embodiments, c is 7. In some embodiments, c is 8. In some embodiments, c is 9. In some embodiments, c is 10.

[0237] In some embodiments, d is 1-10. In some embodiments, d is 2-10. In some embodiments, d is 1-5. In some embodiments, d is 2-5. In some embodiments, d is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, d is an integer selected from 2, 3, 4, or 5. In some embodiments, d is 1. In some embodiments, d is 2. In some embodiments, d is 3. In some embodiments, d is 4. In some embodiments, dis 5. In some embodiments, d is 6. In some embodiments, d is 7. In some embodiments, d is 8. In some embodiments, d is 9. In some embodiments, d is 10.

[0238] In some embodiments, e is 1-10. In some embodiments, e is 2-10. In some embodiments, e is 1-5. In some embodiments, e is 2-5. In some embodiments, e is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, e is an integer selected from 2, 3, 4, or 5. In some embodiments, e is 1. In some embodiments, e is 2. In some embodiments, e is 3. In some embodiments, e is 4. In some embodiments, e is 5. In some embodiments, e is 6. In some embodiments, e is 7. In some embodiments, e is 8. In some embodiments, e is 9. In some embodiments, e is 10.

[0239] In some embodiments, g is 1-10. In some embodiments, g is 2-10. In some embodiments, g is 1-5. In some embodiments, g is 2-5. In some embodiments, g is an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, g is an integer selected from 2, 3, 4, 5, or 6. In some embodiments, g is 1. In some embodiments, g is 2. In some embodiments, g is 3. In some embodiments, g is 4. In some embodiments, g is 5. In some embodiments, g is 6. In some embodiments, g is 7. In some embodiments, g is 8. In some embodiments, g is 9. In some embodiments, g is 10.

[0240] In some embodiments, h is 1-10. In some embodiments, h is 2-10. In some embodiments, h is 1-5. In some embodiments, h is 2-5. In some embodiments, h is an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, h is an integer selected from 2, 3, 4, 5, or 6. In some embodiments, h is 1. In some embodiments, h is 2. In some embodiments, h is 3. In some embodiments, h is 4. In some embodiments, h is 5. In some embodiments, h is 6. In some embodiments, h is 7. In some embodiments, h is 8. In some embodiments, h is 9. In some embodiments, h is 10.

[0241] In some embodiments, i is 1-10. In some embodiments, i is 2-10. In some embodiments, i is 1-5. In some embodiments, i is 2-5. In some embodiments, i is an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, i is an integer selected from 2, 3, 4, 5, or 6. In some embodiments, i is 1. In some embodiments, i is 2. In some embodiments, i is 3. In some embodiments, i is 4. In some embodiments, i is 5. In some embodiments, i is 6. In some embodiments, i is 7. In some embodiments, iAttorney Docket No.: 2010794-3211is 8. In some embodiments, i is 9. In some embodiments, i is 10.

[0242] In some embodiments, j is 1-10. In some embodiments, j is 2-10. In some embodiments, j is 1-5. In some embodiments, j is 2-5. In some embodiments, j is an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, j is an integer selected from 2, 3, 4, 5, or 6. In some embodiments, j is 1. In some embodiments, j is 2. In some embodiments, j is 3. In some embodiments, j is 4. In some embodiments, j is 5. In some embodiments, j is 6. In some embodiments, j is 7. In some embodiments, j is 8. In some embodiments,] is 9. In some embodiments,] is 10.

[0243] In some embodiments, a sum of a, b, c, d, and e is not greater than 30. In some embodiments, a sum of a, b, c, d, and e is not greater than 25. In some embodiments, a sum of a, b, c, d, and e is not greater than 24. In some embodiments, a sum of a, b, c, d, and e is not greater than 23. In some embodiments, a sum of a, b, c, d, and e is not greater than 22. In some embodiments, a sum of a, b, c, d, and e is not greater than 21. In some embodiments, a sum of a, b, c, d, and e is not greater than 20. In some embodiments, a sum of a, b, c, d, and e is not greater than 19. In some embodiments, a sum of a, b, c, d, and e is not greater than 18. In some embodiments, a sum of a, b, c, d, and e is not greater than 17. In some embodiments, a sum of a, b, c, d, and e is not greater than 16. In some embodiments, a sum of a, b, c, d, and e is not greater than 15. In some embodiments, a sum of a, b, c, d, and e is not greater than 14. In some embodiments, a sum of a, b, c, d, and e is not greater than 13. In some embodiments, a sum of a, b, c, d, and e is not greater than 12. In some embodiments, a sum of a, b, c, d, and e is not greater than 11. In some embodiments, a sum of a, b, c, d, and e is not greater than 10.

[0244] In some embodiments, a sum of g, h, i, and j is not greater than 30. In some embodiments, a sum of g, h, i, and j is not greater than 25. In some embodiments, a sum of g, h, i, and j is not greater than 24. In some embodiments, a sum of g, h, i, and j is not greater than 23. In some embodiments, a sum of g, h, i, and j is not greater than 22. In some embodiments, a sum of g, h, i, and j is not greater than 21. In some embodiments, a sum of g, h, i, and j is not greater than 20. In some embodiments, a sum of g, h, i, and j is not greater than 19. In some embodiments, a sum of g, h, i, and j is not greater than 18. In some embodiments, a sum of g, h, i, and j is not greater than 17. In some embodiments, a sum of g, h, i, and j is not greater than 16. In some embodiments, a sum of g, h, i, and j is not greater than 15. In some embodiments, a sum of g, h, i, and j is not greater than 14. In some embodiments, a sum of g, h, i, and j is not greater than 13. In some embodiments, a sum of g, h, i, and j is not greater than 12. In some embodiments, a sum of g, h, i, and j is not greater than 11. In some embodiments, a sum of g, h, i, and j is not greater than 10.

[0245] In some embodiments, each of a, b, c, d, and e is independently 1-10. In some embodiments, each of a, b, c, d, and e is independently 2-10. In some embodiments, each of a, b, c, d, and e is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of b, c, d, and e isAttorney Docket No.: 2010794-3211independently an integer selected from 2, 3, 4, or 5. In some embodiments, each of b, c, d, and e is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, d, and e is independently an integer selected from 2, 3, 4, or 5. In some embodiments, each of b, d, and e is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of b, d, and e is independently an integer selected from 2, 3, 4, or 5.

[0246] In some embodiments, each of g, h, i, and j is independently 1-10. In some embodiments, each of g, h, i, and j is independentiy 2-10. In some embodiments, each of g, h, i, and j is independently an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, each of g, h, i, and j is independently an integer selected from 2, 3, 4, 5, or 6.

[0247] In some embodiments, a is 3, b is 2, c is 4, d is 3, and e is 3. In some embodiments, a is 1, b is 2, c is 4, d is 3, and e is 3. In some embodiments, g is 3, h is 3, i is 3, and j is 6. In some embodiments, a is 3, b is 2, c is 4, d is 3, e is 3, g is 3, h is 3, i is 3, and j is 6. In some embodiments, a is 1, b is 2, c is 4, d is 3, e is 3, g is 3, h is 3, i is 3, and j is 6.

[0248] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN and a sense strand comprising fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN and a sense strand that is fN mN fN mN mN mN fN mN fN f fN mN mN mN fN mN mN mN mN mN mN.

[0249] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN and a sense strand comprising f mN fN mN mN mN fN mN fN fN fN mN mN mN f mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN f mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN and a sense strand that is fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN.

[0250] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN and a sense strand comprising fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN and a sense strand that is fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0251] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fNAttorney Docket No.: 2010794-3211* mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN and a sense strand comprising fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN and a sense strand that is fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0252] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN and a sense strand comprising fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN and a sense strand that is fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0253] In some embodiments, a pattern of sugar modifications comprises mN fN (mN), xN yN xN (yN)b (mN)c fN mN fN (mN)d fN (mN),, wherein N represents any nucleoside; m represents a 2’-0Me modification; f represents a 2’ -F modification; xN represents a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-0Me modified nucleoside; yN represents GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-F modified nucleoside; and each of a, b, c, d, and e are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN), xN yN xN (yN)b (mN), fN mN fN (mN)d fN (mN)e, wherein N represents any nucleoside; m represents a 2’-0Me modification; f represents a 2’-F modification; xN represents a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-OMe modified nucleoside; yN represents GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-F modified nucleoside; and each of a, b, c, d, and e are independently an integer of 1 or greater.

[0254] In some embodiments, an oligonucleotide comprises mN fN (mN), xN yN xN (yN)b (mN)cfN mN fN (mN)d fN (mN)e, wherein N represents any nucleoside; m represents a 2’-0Me modification; f represents a 2’-F modification; xN represents a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-OMe modified nucleoside; yN represents GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-F modified nucleoside; and each of a, b, c, d, and e are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN), xN yN xN (yN)b (mN), fN mN fN (mN)d fN (mN),, wherein N represents any nucleoside; m represents a 2’-0Me modification; f represents a 2’-F modification; xN represents a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-0Me modified nucleoside; yN represents GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-F modified nucleoside; andAttorney Docket No.: 2010794-3211each of a, b, c, d, and e are independently an integer of 1 or greater. In some embodiments, an antisense strand comprises mN fN (mN)axN yN xN (yN)b (mN)cfN mN fN (mN)d fN (mN)e, wherein N represents any nucleoside; m represents a 2’-0Me modification; f represents a 2’-F modification; xN represents a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-0Me modified nucleoside; yN represents GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-F modified nucleoside; and each of a, b, c, d, and e are independently an integer of 1 or greater. In some embodiments, an antisense strand is mN fN (mN), xN yN xN (yN)b (mN)cfN mN fN (mN)d fN (mN)e, wherein N represents any nucleoside; m represents a 2’-0Me modification; f represents a 2’-F modification; xN represents a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-0Me modified nucleoside; yN represents GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-F modified nucleoside; and each of a, b, c, d, and e are independently an integer of 1 or greater.

[0255] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN), xN yN xN (yN)b (mN)cfN mN fN (mN)d f (mN)e, wherein N represents any nucleoside; m represents a 2’-0Me modification; f represents a 2’-F modification; xN represents a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-OMe modified nucleoside; yN represents GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-F modified nucleoside; and each of a, b, c, d, and e are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN), xN yN xN (yN)b (mN)cfN mN fN (mN)d fN (mN)e, wherein N represents any nucleoside; m represents a 2’-OMe modification; f represents a 2’-F modification; xN represents a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-0Me modified nucleoside; yN represents GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-F modified nucleoside; and each of a, b, c, d, and e are independenfly an integer of 1 or greater.

[0256] In some embodiments, a is 1-10. In some embodiments, a is 2-10. In some embodiments, a is 1-5. In some embodiments, a is 2-5. In some embodiments, a is an integer selected from 2, 3, 4, 5, 6, or 7. In some embodiments, a is an integer selected from 2, 3, 4, or 5. In some embodiments, a is 1. In some embodiments, a is 2. In some embodiments, a is 3. In some embodiments, a is 4. In some embodiments, a is 5. In some embodiments, a is 6. In some embodiments, a is 7. In some embodiments, a is 8. In some embodiments, a is 9. In some embodiments, a is 10.

[0257] In some embodiments, b is 1-10. In some embodiments, b is 2-10. In some embodiments, b is 1-5. In some embodiments, b is 2-5. In some embodiments, b is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, b is an integer selected from 2, 3, 4, or 5. In some embodiments, In some embodiments, b is 1. In some embodiments, b is 2. In some embodiments, b is 3. In some embodiments, b is 4. In some embodiments, b is 5. In some embodiments, b is 6. In some embodiments, b is 7. In some embodiments, b is 8. In some embodiments, b is 9. In some embodiments, b is 10.Attorney Docket No.: 2010794-3211

[0258] In some embodiments, c is 1-10. In some embodiments, c is 2-10. In some embodiments, c is 1-5. In some embodiments, c is 2-5. In some embodiments, c is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, c is an integer selected from 2, 3, 4, or 5. In some embodiments, c is 1. In some embodiments, c is 2. In some embodiments, c is 3. In some embodiments, c is 4. In some embodiments, c is 5. In some embodiments, c is 6. In some embodiments, c is 7. In some embodiments, c is 8. In some embodiments, c is 9. In some embodiments, c is 10.

[0259] In some embodiments, d is 1-10. In some embodiments, d is 2-10. In some embodiments, d is 1-5. In some embodiments, d is 2-5. In some embodiments, d is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, d is an integer selected from 2, 3, 4, or 5. In some embodiments, d is 1. In some embodiments, d is 2. In some embodiments, d is 3. In some embodiments, d is 4. In some embodiments, dis 5. In some embodiments, d is 6. In some embodiments, d is 7. In some embodiments, d is 8. In some embodiments, d is 9. In some embodiments, d is 10.

[0260] In some embodiments, e is 1-10. In some embodiments, e is 2-10. In some embodiments, e is 1-5. In some embodiments, e is 2-5. In some embodiments, e is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, e is an integer selected from 2, 3, 4, or 5. In some embodiments, e is 1. In some embodiments, e is 2. In some embodiments, e is 3. In some embodiments, e is 4. In some embodiments, e is 5. In some embodiments, e is 6. In some embodiments, e is 7. In some embodiments, e is 8. In some embodiments, e is 9. In some embodiments, e is 10.

[0261] In some embodiments, a sum of a, b, c, d, and e is not greater than 30. In some embodiments, a sum of a, b, c, d, and e is not greater than 25. In some embodiments, a sum of a, b, c, d, and e is not greater than 24. In some embodiments, a sum of a, b, c, d, and e is not greater than 23. In some embodiments, a sum of a, b, c, d, and e is not greater than 22. In some embodiments, a sum of a, b, c, d, and e is not greater than 21. In some embodiments, a sum of a, b, c, d, and e is not greater than 20. In some embodiments, a sum of a, b, c, d, and e is not greater than 19. In some embodiments, a sum of a, b, c, d, and e is not greater than 18. In some embodiments, a sum of a, b, c, d, and e is not greater than 17. In some embodiments, a sum of a, b, c, d, and e is not greater than 16. In some embodiments, a sum of a, b, c, d, and e is not greater than 15. In some embodiments, a sum of a, b, c, d, and e is not greater than 14. In some embodiments, a sum of a, b, c, d, and e is not greater than 13. In some embodiments, a sum of a, b, c, d, and e is not greater than 12. In some embodiments, a sum of a, b, c, d, and e is not greater than 11. In some embodiments, a sum of a, b, c, d, and e is not greater than 10.

[0262] In some embodiments, each of a, b, c, d, and e is independently 1-10. In some embodiments, each of a, b, c, d, and e is independently 2-10. In some embodiments, each of a, b, c, d, and e is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of b, c, d, and e is independently an integer selected from 2, 3, 4, or 5. In some embodiments, each of b, c, d, and e isAttorney Docket No.: 2010794-3211independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, d, and e is independently an integer selected from 2, 3, 4, or 5. In some embodiments, each of b, d, and e is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of b, d, and e is independently an integer selected from 2, 3, 4, or 5.

[0263] In some embodiments, a is 2, b is 2, c is 4, d is 3, and e is 3.

[0264] In some embodiments, each xN is independently a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-OMe modified nucleoside. In some embodiments, at least one xN is a GNA nucleoside. In some embodiments, at least one xN is a UNA nucleoside. In some embodiments, at least one xN is a DNA nucleoside. In some embodiments, at least one xN is a 2’-OMe modified nucleoside. In some embodiments, at least 2 xN are independently GNA nucleosides. In some embodiments, at least 2 xN are independently UNA nucleosides. In some embodiments, at least 2 xN are independently DNA nucleosides. In some embodiments, at least 2 xN are independently 2’-OMe modified nucleosides.

[0265] In some embodiments, each yN is independently a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’F modified nucleoside. In some embodiments, at least one yN is a GNA nucleoside. In some embodiments, at least one yN is a UNA nucleoside. In some embodiments, at least one yN is a DNA nucleoside. In some embodiments, at least one yN is a 2’-F modified nucleoside. In some embodiments, at least 2 yN are independently GNA nucleosides. In some embodiments, at least 2 yN are independently UNA nucleosides. In some embodiments, at least 2 yN are independently DNA nucleosides. In some embodiments, at least 2 yN are independently 2’-F modified nucleosides.

[0266] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

[0267] In some embodiments, an oligonucleotide comprises mN fN mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an oligonucleotide is mN fN mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

[0268] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

[0269] In some embodiments, a pattern of sugar modifications comprises mN * fN * mN mN xN yNAttorney Docket No.: 2010794-3211xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN * mN* mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, a pattern of sugar modifications is mN * fN * mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0270] In some embodiments, an oligonucleotide comprises mN * fN * mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense stand comprises mN * fN * mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * fN * mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0271] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense stand that is mN * fN * mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0272] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

[0273] In some embodiments, an oligonucleotide comprises mN fN mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an oligonucleotide is mN fN mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

[0274] In some embodiments, an oligonucleotide comprises an antisense stand comprising mN fN mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

[0275] In some embodiments, a pattern of sugar modifications comprises mN * fN * mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents aAttorney Docket No.: 2010794-3211phosphorothioate internucleotidic linkage. In some embodiments, a pattern of sugar modifications is mN * fN * mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0276] In some embodiments, an oligonucleotide comprises mN - f * mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * f * mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0277] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0278] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN), xN yN xN (yN)b (mN)cfN mN fN (mN)d fN (mN)eand a sense strand comprising fN mN fN (mN)gfN mN (fN)h (mN)i fN (mN)j, wherein N represents any nucleoside; m represents a 2’-0Me modification; f represents a 2’-F modification; xN represents a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-0Me modified nucleoside; yN represents GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-F modified nucleoside; and each of a, b, c, d, e, g, h, i, and j are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN), xN yN xN (yN)b (mN)cfN mN fN (mN)d fN (mN)eand a sense strand that is fN mN fN (mN). fN mN (fN)h (mN)i fN (mN)j, wherein N represents any nucleoside; m represents a 2’-0Me modification; f represents a 2’-F modification; xN represents a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-0Me modified nucleoside; yN represents GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’-F modified nucleoside; and each of a, b, c, d, e, g, h, i, and j are independently an integer of 1 or greater.

[0279] In some embodiments, a is 1-10. In some embodiments, a is 2-10. In some embodiments, a is 1-5. In some embodiments, a is 2-5. In some embodiments, a is an integer selected from 2, 3, 4, 5, 6, or 7. In some embodiments, a is an integer selected from 2, 3, 4, or 5. In some embodiments, a is 1. In some embodiments, a is 2. In some embodiments, a is 3. In some embodiments, a is 4. In some embodiments, a is 5. In some embodiments, a is 6. In some embodiments, a is 7. In some embodiments, a is 8. In someAttorney Docket No.: 2010794-3211embodiments, a is 9. In some embodiments, a is 10.

[0280] In some embodiments, b is 1-10. In some embodiments, b is 2-10. In some embodiments, b is 1-5. In some embodiments, b is 2-5. In some embodiments, b is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, b is an integer selected from 2, 3, 4, or 5. In some embodiments, b is 1. In some embodiments, b is 2. In some embodiments, b is 3. In some embodiments, b is 4. In some embodiments, b is 5. In some embodiments, b is 6. In some embodiments, b is 7. In some embodiments, b is 8. In some embodiments, b is 9. In some embodiments, b is 10.

[0281] In some embodiments, c is 1-10. In some embodiments, c is 2-10. In some embodiments, c is 1-5. In some embodiments, c is 2-5. In some embodiments, c is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, c is an integer selected from 2, 3, 4, or 5. In some embodiments, c is 1. In some embodiments, c is 2. In some embodiments, c is 3. In some embodiments, c is 4. In some embodiments, c is 5. In some embodiments, c is 6. In some embodiments, c is 7. In some embodiments, c is 8. In some embodiments, c is 9. In some embodiments, c is 10.

[0282] In some embodiments, d is 1-10. In some embodiments, d is 2-10. In some embodiments, d is 1-5. In some embodiments, d is 2-5. In some embodiments, d is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, d is an integer selected from 2, 3, 4, or 5. In some embodiments, d is 1. In some embodiments, d is 2. In some embodiments, d is 3. In some embodiments, d is 4. In some embodiments, dis 5. In some embodiments, d is 6. In some embodiments, d is 7. In some embodiments, d is 8. In some embodiments, d is 9. In some embodiments, d is 10.

[0283] In some embodiments, e is 1-10. In some embodiments, e is 2-10. In some embodiments, e is 1-5. In some embodiments, e is 2-5. In some embodiments, e is an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, e is an integer selected from 2, 3, 4, or 5. In some embodiments, e is 1. In some embodiments, e is 2. In some embodiments, e is 3. In some embodiments, e is 4. In some embodiments, e is 5. In some embodiments, e is 6. In some embodiments, e is 7. In some embodiments, e is 8. In some embodiments, e is 9. In some embodiments, e is 10.

[0284] In some embodiments, g is 1-10. In some embodiments, g is 2-10. In some embodiments, g is 1-5. In some embodiments, g is 2-5. In some embodiments, g is an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, g is an integer selected from 2, 3, 4, 5, or 6. In some embodiments, g is 1. In some embodiments, g is 2. In some embodiments, g is 3. In some embodiments, g is 4. In some embodiments, g is 5. In some embodiments, g is 6. In some embodiments, g is 7. In some embodiments, g is 8. In some embodiments, g is 9. In some embodiments, g is 10.

[0285] In some embodiments, h is 1-10. In some embodiments, h is 2-10. In some embodiments, h is 1-5. In some embodiments, h is 2-5. In some embodiments, h is an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, h is an integer selected from 2, 3, 4, 5, or 6. In some embodiments, h is 1. InAttorney Docket No.: 2010794-3211some embodiments, h is 2. In some embodiments, h is 3. In some embodiments, h is 4. In some embodiments, h is 5. In some embodiments, h is 6. In some embodiments, h is 7. In some embodiments, h is 8. In some embodiments, h is 9. In some embodiments, h is 10.

[0286] In some embodiments, i is 1-10. In some embodiments, i is 2-10. In some embodiments, i is 1-5. In some embodiments, i is 2-5. In some embodiments, i is an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, i is an integer selected from 2, 3, 4, 5, or 6. In some embodiments, i is 1. In some embodiments, i is 2. In some embodiments, i is 3. In some embodiments, i is 4. In some embodiments, i is 5. In some embodiments, i is 6. In some embodiments, i is 7. In some embodiments, i is 8. In some embodiments, i is 9. In some embodiments, i is 10.

[0287] In some embodiments, j is 1-10. In some embodiments, j is 2-10. In some embodiments, j is 1-5. In some embodiments, j is 2-5. In some embodiments, j is an integer selected from 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, j is an integer selected from 2, 3, 4, 5, or 6. In some embodiments, j is 1. In some embodiments, j is 2. In some embodiments, j is 3. In some embodiments, j is 4. In some embodiments, j is 5. In some embodiments, j is 6. In some embodiments, j is 7. In some embodiments, j is 8. In some embodiments,] is 9. In some embodiments,] is 10.

[0288] In some embodiments, a sum of a, b, c, d, and e is not greater than 30. In some embodiments, a sum of a, b, c, d, and e is not greater than 25. In some embodiments, a sum of a, b, c, d, and e is not greater than 24. In some embodiments, a sum of a, b, c, d, and e is not greater than 23. In some embodiments, a sum of a, b, c, d, and e is not greater than 22. In some embodiments, a sum of a, b, c, d, and e is not greater than 21. In some embodiments, a sum of a, b, c, d, and e is not greater than 20. In some embodiments, a sum of a, b, c, d, and e is not greater than 19. In some embodiments, a sum of a, b, c, d, and e is not greater than 18. In some embodiments, a sum of a, b, c, d, and e is not greater than 17. In some embodiments, a sum of a, b, c, d, and e is not greater than 16. In some embodiments, a sum of a, b, c, d, and e is not greater than 15. In some embodiments, a sum of a, b, c, d, and e is not greater than 14. In some embodiments, a sum of a, b, c, d, and e is not greater than 13. In some embodiments, a sum of a, b, c, d, and e is not greater than 12. In some embodiments, a sum of a, b, c, d, and e is not greater than 11. In some embodiments, a sum of a, b, c, d, and e is not greater than 10.

[0289] In some embodiments, a sum of g, h, i, and j is not greater than 30. In some embodiments, a sum of g, h, i, and j is not greater than 25. In some embodiments, a sum of g, h, i, and j is not greater than 24. In some embodiments, a sum of g, h, i, and j is not greater than 23. In some embodiments, a sum of g, h, i, and j is not greater than 22. In some embodiments, a sum of g, h, i, and j is not greater than 21. In some embodiments, a sum of g, h, i, and j is not greater than 20. In some embodiments, a sum of g, h, i, and j is not greater than 19. In some embodiments, a sum of g, h, i, and j is not greater than 18. In some embodiments, a sum of g, h, i, and j is not greater than 17. In some embodiments, a sum of g, h, i, and j isAttorney Docket No.: 2010794-3211not greater than 16. In some embodiments, a sum of g, h, i, and j is not greater than 15. In some embodiments, a sum of g, h, i, and j is not greater than 14. In some embodiments, a sum of g, h, i, and j is not greater than 13. In some embodiments, a sum of g, h, i, and j is not greater than 12. In some embodiments, a sum of g, h, i, and j is not greater than 11. In some embodiments, a sum of g, h, i, and j is not greater than 10.

[0290] In some embodiments, each of a, b, c, d, and e is independently 1-10. In some embodiments, each of a, b, c, d, and e is independently 2-10. In some embodiments, each of a, b, c, d, and e is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of b, c, d, and e is independently an integer selected from 2, 3, 4, or 5. In some embodiments, each of b, c, d, and e is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, d, and e is independently an integer selected from 2, 3, 4, or 5. In some embodiments, each of b, d, and e is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of b, d, and e is independently an integer selected from 2, 3, 4, or 5.

[0291] In some embodiments, each of g, h, i, and j is independently 1-10. In some embodiments, each of g, h, i, and j is independently 2-10. In some embodiments, each of g, h, i, and j is independently an integer selected from 2, 3, 4, 5, 6, 7. 8, or 9. In some embodiments, each of g, h, i, and j is independently an integer selected from 2, 3, 4, 5, or 6.

[0292] In some embodiments, a is 2, b is 2, c is 4, d is 3, and e is 3. In some embodiments, g is 3, h is 3, i is 3, and j is 6. In some embodiments, a is 2, b is 2, c is 4, d is 3, e is 3, g is 3, h is 3, i is 3, and j is 6.

[0293] In some embodiments, each xN is independently a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’ -OMe modified nucleoside. In some embodiments, at least one xN is a GNA nucleoside. In some embodiments, at least one xN is a UNA nucleoside. In some embodiments, at least one xN is a DNA nucleoside. In some embodiments, at least one xN is a 2’ -OMe modified nucleoside. In some embodiments, at least 2 xN are independently GNA nucleosides. In some embodiments, at least 2 xN are independently UNA nucleosides. In some embodiments, at least 2 xN are independently DNA nucleosides. In some embodiments, at least 2 xN are independently 2’ -OMe modified nucleosides.

[0294] In some embodiments, each yN is independently a GNA nucleoside, a UNA nucleoside, a DNA nucleoside, or a 2’F modified nucleoside. In some embodiments, at least one yN is a GNA nucleoside. In some embodiments, at least one yN is a UNA nucleoside. In some embodiments, at least one yN is a DNA nucleoside. In some embodiments, at least one yN is a 2’-F modified nucleoside. In some embodiments, at least 2 yN are independently GNA nucleosides. In some embodiments, at least 2 yN are independently UNA nucleosides. In some embodiments, at least 2 yN are independently DNA nucleosides. In some embodiments, at least 2 yN are independently 2’-F modified nucleosides.

[0295] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fNAttorney Docket No.: 2010794-3211mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN mN mN, and a sense strand comprising fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN mN mN, and a sense strand that is fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN.

[0296] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN f mN mN mN, and a sense strand comprising fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN, and a sense strand that is fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN.

[0297] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, and a sense strand comprising fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN xN yN xN yN yN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, and a sense strand that is fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0298] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, and a sense strand comprising fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN xN yN xN yN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, and a sense strand that is fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0299] In some embodiments, a pattern of sugar modifications comprises mN fN (mN), fN mN (fN)b (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN)afN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of a, b, c, and d are independently an integer of 1 or greater.

[0300] In some embodiments, an oligonucleotide comprises mN fN (mN), fN mN (fN)b (mN)cfN mNAttorney Docket No.: 2010794-3211fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN)afN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand comprises mN fN (mN), fN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand is mN fN (mN)afN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, and each of a, b, c, and d are independently an integer of 1 or greater.

[0301] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN), fN mN (fN)b(mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN)afN mN (fN)b (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, and each of a, b, c, and d are independently an integer of 1 or greater.

[0302] In some embodiments, each of a, b, c, and d is as described herein.

[0303] In some embodiments, a sum of a, b, c, and d is not greater than 30. In some embodiments, a sum of a, b, c, and d is not greater than 25. In some embodiments, a sum of a, b, c, and d is not greater than 24. In some embodiments, a sum of a, b, c, and d is not greater than 23. In some embodiments, a sum of a, b, c, and d is not greater than 22. In some embodiments, a sum of a, b, c, and d is not greater than 21. In some embodiments, a sum of a, b, c, and d is not greater than 20. In some embodiments, a sum of a, b, c, and d is not greater than 19. In some embodiments, a sum of a, b, c, and d is not greater than 18. In some embodiments, a sum of a, b, c, and d is not greater than 17. In some embodiments, a sum of a, b, c, and d is not greater than 16. In some embodiments, a sum of a, b, c, and d is not greater than 15. In some embodiments, a sum of a, b, c, and d is not greater than 14. In some embodiments, a sum of a, b, c, and d is not greater than 13. In some embodiments, a sum of a, b, c, and d is not greater than 12. In some embodiments, a sum of a, b, c, and d is not greater than 11. In some embodiments, a sum of a, b, c, and d is not greater than 10.

[0304] In some embodiments, each of a, b, c, and d is independently 1-10. In some embodiments, each of a, b, c, and d is independently 2-10. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, or 7.Attomey Docket No.: 2010794-3211

[0305] In some embodiments, a is 3, b is 2, c is 4, and d is 7.

[0306] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0307] In some embodiments, an oligonucleotide comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0308] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN f mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0309] In some embodiments, an oligonucleotide comprises mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0310] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * IN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0311] In some embodiments, a pattern of sugar modifications comprises mN fN (mN)agN fN mN (fN)b (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN), gN fN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-Attorney Docket No.: 2010794-3211OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0312] In some embodiments, an oligonucleotide comprises mN fN (mN)agN fN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’ -OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN), gN fN mN (fN)t (mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense stand comprises mN fN (mN), gN fN mN (fN)t (mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’ -OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense stand is mN fN (mN)agN fN mN (fN)t (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0313] In some embodiments, an oligonucleotide comprises an antisense stand comprising mN fN (mN), gN fN mN (fN)b (mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense stand that is mN fN (mN), gN fN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’ -OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0314] In some embodiments, each of a, b, c, and d is as described herein.

[0315] In some embodiments, a sum of a, b, c, and d is not greater than 30. In some embodiments, a sum of a, b, c, and d is not greater than 25. In some embodiments, a sum of a, b, c, and d is not greater than 24. In some embodiments, a sum of a, b, c, and d is not greater than 23. In some embodiments, a sum of a, b, c, and d is not greater than 22. In some embodiments, a sum of a, b, c, and d is not greater than 21. In some embodiments, a sum of a, b, c, and d is not greater than 20. In some embodiments, a sum of a, b, c, and d is not greater than 19. In some embodiments, a sum of a, b, c, and d is not greater than 18. In some embodiments, a sum of a, b, c, and d is not greater than 17. In some embodiments, a sum of a, b, c, and d is not greater than 16. In some embodiments, a sum of a, b, c, and d is not greater than 1 . In some embodiments, a sum of a, b, c, and d is not greater than 14. In some embodiments, a sum of a, b, c, and d is not greater than 13. In some embodiments, a sum of a, b, c, and d is not greater than 12. In some embodiments, a sum of a, b, c, and d is not greater than 11. In some embodiments, a sum of a, b, c, and dAttorney Docket No.: 2010794-3211is not greater than 10.

[0316] In some embodiments, each of a, b, c, and d is independently 1-10. In some embodiments, each of a, b, c, and d is independently 2-10. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, or 7.

[0317] In some embodiments, a is 2, b is 2, c is 4, and d is 7.

[0318] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0319] In some embodiments, an oligonucleotide comprises mN fN mN mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide is mN fN mN mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0320] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0321] In some embodiments, an oligonucleotide comprises mN * fN * mN mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN mN gN fN mN fN IN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * f * mN mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0322] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN mN * mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN gN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.Attorney Docket No.: 2010794-3211

[0323] In some embodiments, a pattern of sugar modifications comprises mN fN (mN), gN mN (fN)b(mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN), gN mN (fN)b (mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0324] In some embodiments, an oligonucleotide comprises mN fN (mN), gN mN (fN)b(mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN), gN mN (fN)b(mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand comprises mN fN (mN), gN mN (fN)b(mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand is mN fN (mN), gN mN (fN)b(mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0325] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN), gN mN (fN)b(mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN), gN mN (fN)b(mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0326] In some embodiments, each of a, b, c, and d is as described herein.

[0327] In some embodiments, a sum of a, b, c, and d is not greater than 30. In some embodiments, a sum of a, b, c, and d is not greater than 25. In some embodiments, a sum of a, b, c, and d is not greater than 24. In some embodiments, a sum of a, b, c, and d is not greater than 23. In some embodiments, a sum of a, b, c, and d is not greater than 22. In some embodiments, a sum of a, b, c, and d is not greater than 21. In some embodiments, a sum of a, b, c, and d is not greater than 20. In some embodiments, a sum of a, b, c, and d is not greater than 19. In some embodiments, a sum of a, b, c, and d is not greater than 18. In someAttorney Docket No.: 2010794-3211embodiments, a sum of a, b, c, and d is not greater than 17. In some embodiments, a sum of a, b, c, and d is not greater than 16. In some embodiments, a sum of a, b, c, and d is not greater than 15. In some embodiments, a sum of a, b, c, and d is not greater than 14. In some embodiments, a sum of a, b, c, and d is not greater than 13. In some embodiments, a sum of a, b, c, and d is not greater than 12. In some embodiments, a sum of a, b, c, and d is not greater than 11. In some embodiments, a sum of a, b, c, and d is not greater than 10.

[0328] In some embodiments, each of a, b, c, and d is independently 1-10. In some embodiments, each of a, b, c, and d is independently 2-10. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, and d is independently an integer selected from 2. 3, 4, 5, 6, or 7.

[0329] In some embodiments, a is 3, b is 2, c is 4, and d is 7.

[0330] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0331] In some embodiments, an oligonucleotide comprises mN fN mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide is mN fN mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0332] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN IN mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0333] In some embodiments, an oligonucleotide comprises mN * fN * mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * fN * mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.Attorney Docket No.: 2010794-3211

[0334] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN mN gN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0335] In some embodiments, a pattern of sugar modifications comprises mN fN (mN), fN gN (fN)b (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN), fN gN (fN)b (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0336] In some embodiments, an oligonucleotide comprises mN fN (mN), fN gN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN), fN gN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand comprises mN fN (mN), fN gN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand is mN fN (mN), fN gN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0337] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN), fN gN (fN)b (mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN), fN gN (fN)b(mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0338] In some embodiments, each of a, b, c, and d is as described herein.

[0339] In some embodiments, a sum of a, b, c, and d is not greater than 30. In some embodiments, a sum of a, b, c, and d is not greater than 25. In some embodiments, a sum of a, b, c, and d is not greater thanAttorney Docket No.: 2010794-321124. In some embodiments, a sum of a, b, c, and d is not greater than 23. In some embodiments, a sum of a, b, c, and d is not greater than 22. In some embodiments, a sum of a, b, c, and d is not greater than 21. In some embodiments, a sum of a, b, c, and d is not greater than 20. In some embodiments, a sum of a, b, c, and d is not greater than 19. In some embodiments, a sum of a, b, c, and d is not greater than 18. In some embodiments, a sum of a, b, c, and d is not greater than 17. In some embodiments, a sum of a, b, c, and d is not greater than 16. In some embodiments, a sum of a, b, c, and d is not greater than 15. In some embodiments, a sum of a, b, c, and d is not greater than 14. In some embodiments, a sum of a, b, c, and d is not greater than 13. In some embodiments, a sum of a, b, c, and d is not greater than 12. In some embodiments, a sum of a, b, c, and d is not greater than 11. In some embodiments, a sum of a, b, c, and d is not greater than 10.

[0340] In some embodiments, each of a, b, c, and d is independently 1-10. In some embodiments, each of a, b, c, and d is independently 2-10. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, or 7.

[0341] In some embodiments, a is 3, b is 2, c is 4, and d is 7.

[0342] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0343] In some embodiments, an oligonucleotide comprises mN fN mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide is mN fN mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0344] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0345] In some embodiments, an oligonucleotide comprises mN * fN * mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN mN fNAttorney Docket No.: 2010794-3211gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * fN * mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0346] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN mN fN gN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0347] In some embodiments, a pattern of sugar modifications comprises mN fN (mN)afN mN gN fN (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, c, and d are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN)afN mN gN fN (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, c, and d are independently an integer of 1 or greater.

[0348] In some embodiments, an oligonucleotide comprises mN fN (mN), fN mN gN fN (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN)afN mN gN fN (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand comprises mN fN (mN), fN mN gN fN (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand is mN fN (mN)afN mN gN fN (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, c, and d are independently an integer of 1 or greater.

[0349] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN f (mN)afN mN gN fN (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN)afN mN gN fN (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside,Attorney Docket No.: 2010794-3211and each of a, c, and d are independently an integer of 1 or greater.

[0350] In some embodiments, each of a, c, and d is as described herein.

[0351] In some embodiments, a sum of a, c, and d is not greater than 30. In some embodiments, a sum of a, c, and d is not greater than 25. In some embodiments, a sum of a, c, and d is not greater than 24. In some embodiments, a sum of a, c, and d is not greater than 23. In some embodiments, a sum of a, c, and d is not greater than 22. In some embodiments, a sum of a, c, and d is not greater than 21. In some embodiments, a sum of a, c, and d is not greater than 20. In some embodiments, a sum of a, c, and d is not greater than 19. In some embodiments, a sum of a, c, and d is not greater than 18. In some embodiments, a sum of a, c, and d is not greater than 17. In some embodiments, a sum of a, c, and d is not greater than 16. In some embodiments, a sum of a, c, and d is not greater than 15. In some embodiments, a sum of a, c, and d is not greater than 14. In some embodiments, a sum of a, c, and d is not greater than 13. In some embodiments, a sum of a, c, and d is not greater than 12. In some embodiments, a sum of a, c, and d is not greater than 11. In some embodiments, a sum of a, c, and d is not greater than 10.

[0352] In some embodiments, each of a, c, and d is independently 1-10. In some embodiments, each of a, c, and d is independently 2-10. In some embodiments, each of a, c, and d is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, c, and d is independently an integer selected from 2, 3, 4, 5, 6, or 7.

[0353] In some embodiments, a is 3, c is 4, and d is 7.

[0354] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0355] In some embodiments, an oligonucleotide comprises mN fN mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide is mN fN mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0356] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0357] In some embodiments, an oligonucleotide comprises mN * fN * mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioateAttomey Docket No.: 2010794-3211internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * fN * mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0358] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN mN fN mN gN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0359] In some embodiments, a pattern of sugar modifications comprises mN fN (mN)afN mN (fN)h(mN)c gN fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN)afN mN (fN)b (mN)c gN fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0360] In some embodiments, an oligonucleotide comprises mN fN (mN)afN mN (fN)b (mN)cgN fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN), fN mN (fN)b (mN)cgN fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand comprises mN fN (mN)afN mN (fN)b (mN)cgN fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand is mN fN (mN), fN mN (fN)b (mN)cgN fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0361] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN), fN mN (fN)b (mN)cgN fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-Attorney Docket No.: 2010794-3211OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN), fN mN (fN)b (mN)cgN fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’ -OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0362] In some embodiments, each of a, b, c, and d is as described herein.

[0363] In some embodiments, a sum of a, b, c, and d is not greater than 30. In some embodiments, a sum of a, b, c, and d is not greater than 25. In some embodiments, a sum of a, b, c, and d is not greater than 24. In some embodiments, a sum of a, b, c, and d is not greater than 23. In some embodiments, a sum of a, b, c, and d is not greater than 22. In some embodiments, a sum of a, b, c, and d is not greater than 21. In some embodiments, a sum of a, b, c, and d is not greater than 20. In some embodiments, a sum of a, b, c, and d is not greater than 19. In some embodiments, a sum of a, b, c, and d is not greater than 18. In some embodiments, a sum of a, b, c, and d is not greater than 17. In some embodiments, a sum of a, b, c, and d is not greater than 16. In some embodiments, a sum of a, b, c, and d is not greater than 15. In some embodiments, a sum of a, b, c, and d is not greater than 14. In some embodiments, a sum of a, b, c, and d is not greater than 13. In some embodiments, a sum of a, b, c, and d is not greater than 12. In some embodiments, a sum of a, b, c, and d is not greater than 11. In some embodiments, a sum of a, b, c, and d is not greater than 10.

[0364] In some embodiments, each of a, b, c, and d is independently 1-10. In some embodiments, each of a, b, c, and d is independently 2-10. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, and d is independently an integer selected from 2. 3, 4, 5, 6, or 7.

[0365] In some embodiments, a is 3, b is 2, c is 3, and d is 7.

[0366] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN mN mN.

[0367] In some embodiments, an oligonucleotide comprises mN fN mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide is mN f mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN mN mN.

[0368] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fNAttorney Docket No.: 2010794-3211mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN mN mN.

[0369] In some embodiments, an oligonucleotide comprises mN * fN * mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * fN * mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0370] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN mN fN mN fN fN mN mN mN gN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0371] In some embodiments, a pattern of sugar modifications comprises mN fN (mN), fN mN (fN)b (mN), gN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN), fN mN (fN)b (mN), gN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0372] In some embodiments, an oligonucleotide comprises mN fN (mN), fN mN (fN)b (mN), gN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN), fN mN (fN)b (mN), gN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand comprises mN fN (mN), fN mN (fN)b (mN), gN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand is mN fN (mN), fN mN (fN)b (mN), gN mN fNAttorney Docket No.: 2010794-3211(mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0373] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN), fN mN (fN)b (mN)cgN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN), fN mN (fN)b (mN)cgN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0374] In some embodiments, each of a, b, c, and d is as described herein.

[0375] In some embodiments, a sum of a, b, c, and d is not greater than 30. In some embodiments, a sum of a, b, c, and d is not greater than 25. In some embodiments, a sum of a, b, c, and d is not greater than 24. In some embodiments, a sum of a, b, c, and d is not greater than 23. In some embodiments, a sum of a, b, c, and d is not greater than 22. In some embodiments, a sum of a, b, c, and d is not greater than 21. In some embodiments, a sum of a, b, c, and d is not greater than 20. In some embodiments, a sum of a, b, c, and d is not greater than 19. In some embodiments, a sum of a, b, c, and d is not greater than 18. In some embodiments, a sum of a, b, c, and d is not greater than 17. In some embodiments, a sum of a, b, c, and d is not greater than 16. In some embodiments, a sum of a, b, c, and d is not greater than 15. In some embodiments, a sum of a, b, c, and d is not greater than 14. In some embodiments, a sum of a, b, c, and d is not greater than 13. In some embodiments, a sum of a, b, c, and d is not greater than 12. In some embodiments, a sum of a, b, c, and d is not greater than 11. In some embodiments, a sum of a, b, c, and d is not greater than 10.

[0376] In some embodiments, each of a, b, c, and d is independently 1-10. In some embodiments, each of a, b, c, and d is independently 2-10. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, or 7.

[0377] In some embodiments, a is 3, b is 2, c is 4, and d is 7.

[0378] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN mN fN mN IN fN mN mN mN mN gN mN fN mN mN mN mN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN mN fN mN fN fN mN mN mN mN gN mN fN mN mN mN mN mN mN mN.

[0379] In some embodiments, an oligonucleotide comprises mN fN mN mN mN fN mN fN fN mN mN mN mN gN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide is mNAttorney Docket No.: 2010794-3211fN mN mN mN fN mN fN fN mN mN mN mN gN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN mN fN mN fN fN mN mN mN mN gN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN mN fN mN fN fN mN mN mN mN gN mN fN mN mN mN mN mN mN mN.

[0380] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN mN fN mN fN fN mN mN mN mN gN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN mN fN mN fN fN mN mN mN mN gN mN fN mN mN mN mN mN mN mN.

[0381] In some embodiments, an oligonucleotide comprises mN * fN * mN mN mN fN mN fN fN mN mN mN mN gN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN mN mN fN mN fN fN mN mN mN mN gN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN mN fN mN fN fN mN mN mN mN gN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * fN * mN mN mN fN mN fN fN mN mN mN mN gN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0382] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN mN fN mN fN f mN mN mN mN gN mN f mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN mN fN mN fN fN mN mN mN mN gN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0383] In some embodiments, a pattern of sugar modifications comprises mN fN (mN), fN mN (fN)b (mN)cfN gN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN)afN mN (fN)b (mN)c fN gN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0384] In some embodiments, an oligonucleotide comprises mN fN (mN)afN mN (fN)b (mN)cfN gN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN)afN mN (fN)b (mN)cfN gN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification,Attorney Docket No.: 2010794-3211gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand comprises mN fN (mN)afN mN (fN)b (mN)cfN gN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand is mN fN (mN), fN mN (fN)b (mN)cfN gN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0385] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN), fN mN (fN)b (mN)cfN gN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN), fN mN (fN)b (mN)cfN gN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0386] In some embodiments, each of a, b, c, and d is as described herein.

[0387] In some embodiments, a sum of a, b, c, and d is not greater than 30. In some embodiments, a sum of a, b, c, and d is not greater than 25. In some embodiments, a sum of a, b, c, and d is not greater than 24. In some embodiments, a sum of a, b, c, and d is not greater than 23. In some embodiments, a sum of a, b, c, and d is not greater than 22. In some embodiments, a sum of a, b, c, and d is not greater than 21. In some embodiments, a sum of a, b, c, and d is not greater than 20. In some embodiments, a sum of a, b, c, and d is not greater than 19. In some embodiments, a sum of a, b, c, and d is not greater than 18. In some embodiments, a sum of a, b, c, and d is not greater than 17. In some embodiments, a sum of a, b, c, and d is not greater than 16. In some embodiments, a sum of a, b, c, and d is not greater than 15. In some embodiments, a sum of a, b, c, and d is not greater than 14. In some embodiments, a sum of a, b, c, and d is not greater than 13. In some embodiments, a sum of a, b, c, and d is not greater than 12. In some embodiments, a sum of a, b, c, and d is not greater than 11. In some embodiments, a sum of a, b, c, and d is not greater than 10.

[0388] In some embodiments, each of a, b, c, and d is independently 1-10. In some embodiments, each of a, b, c, and d is independently 2-10. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, or 7.

[0389] In some embodiments, a is 3, b is 2, c is 4, and d is 7.

[0390] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN mN mN. In some embodiments, a pattern of sugarAttorney Docket No.: 2010794-3211modifications is mN fN mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN mN mN.

[0391] In some embodiments, an oligonucleotide comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide is mN fN mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN mN mN.

[0392] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN mN mN.

[0393] In some embodiments, an oligonucleotide comprises mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN - fN * mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0394] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN gN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0395] In some embodiments, a pattern of sugar modifications comprises mN fN (mN), fN mN (fN)b (mN)c fN mN gN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification. gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN), fN mN (fN)b (mN)c fN mN gN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0396] In some embodiments, an oligonucleotide comprises mN fN (mN), fN mN (fN)b (mN)cfN mNAttorney Docket No.: 2010794-3211gN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN)afN mN (fN)b (mN)cfN mN gN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand comprises mN fN (mN)afN mN (fN)b (mN)cfN mN gN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand is mN fN (mN), fN mN (fN)b (mN)cfN mN gN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0397] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN), fN mN (fN)b (mN)cfN mN gN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN)afN mN (fN)b (mN)cfN mN gN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, gN represents a GNA nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0398] In some embodiments, each of a, b, c, and d is as described herein.

[0399] In some embodiments, a sum of a, b, c, and d is not greater than 30. In some embodiments, a sum of a, b, c, and d is not greater than 25. In some embodiments, a sum of a, b, c, and d is not greater than 24. In some embodiments, a sum of a, b, c, and d is not greater than 23. In some embodiments, a sum of a, b, c, and d is not greater than 22. In some embodiments, a sum of a, b, c, and d is not greater than 21. In some embodiments, a sum of a, b, c, and d is not greater than 20. In some embodiments, a sum of a, b, c, and d is not greater than 19. In some embodiments, a sum of a, b, c, and d is not greater than 18. In some embodiments, a sum of a, b, c, and d is not greater than 17. In some embodiments, a sum of a, b, c, and d is not greater than 16. In some embodiments, a sum of a, b, c, and d is not greater than 15. In some embodiments, a sum of a, b, c, and d is not greater than 14. In some embodiments, a sum of a, b, c, and d is not greater than 13. In some embodiments, a sum of a, b, c, and d is not greater than 12. In some embodiments, a sum of a, b, c, and d is not greater than 11. In some embodiments, a sum of a, b, c, and d is not greater than 10.

[0400] In some embodiments, each of a, b, c, and d is independently 1-10. In some embodiments, each of a, b, c, and d is independently 2-10. In some embodiments, each of a, b, c, and d is independentlyAttorney Docket No.: 2010794-3211an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, or 7.

[0401] In some embodiments, a is 3, b is 2, c is 4, and d is 7.

[0402] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN mN mN.

[0403] In some embodiments, an oligonucleotide comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN mN mN.

[0404] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN mN mN.

[0405] In some embodiments, an oligonucleotide comprises mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0406] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN gN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0407] In some embodiments, a pattern of sugar modifications comprises mN fN (mN). igN fN mN (fN)b (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d areAttorney Docket No.: 2010794-3211independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN), igN fN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0408] In some embodiments, an oligonucleotide comprises mN fN (mN), igN fN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification. igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN), igN fN mN (fN)b (mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand comprises mN fN (mN), igN fN mN (fN)b (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand is mN fN (mN). igN fN mN (fN)b (mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0409] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN), igN fN mN (fN)b (mN), fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN), igN fN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0410] In some embodiments, each of a, b, c, and d is as described herein.

[0411] In some embodiments, a sum of a, b, c, and d is not greater than 30. In some embodiments, a sum of a, b, c, and d is not greater than 25. In some embodiments, a sum of a, b, c, and d is not greater than 24. In some embodiments, a sum of a, b, c, and d is not greater than 23. In some embodiments, a sum of a, b, c, and d is not greater than 22. In some embodiments, a sum of a, b, c, and d is not greater than 21. In some embodiments, a sum of a, b, c, and d is not greater than 20. In some embodiments, a sum of a, b, c, and d is not greater than 19. In some embodiments, a sum of a, b, c, and d is not greater than 18. In some embodiments, a sum of a, b, c, and d is not greater than 17. In some embodiments, a sum of a, b, c, and d is not greater than 16. In some embodiments, a sum of a, b, c, and d is not greater than 15. In some embodiments, a sum of a, b, c, and d is not greater than 14. In some embodiments, a sum of a, b, c, and dAttorney Docket No.: 2010794-3211is not greater than 13. In some embodiments, a sum of a, b, c, and d is not greater than 12. In some embodiments, a sum of a, b, c, and d is not greater than 11. In some embodiments, a sum of a, b, c, and d is not greater than 10.

[0412] In some embodiments, each of a, b, c, and d is independently 1-10. In some embodiments, each of a, b, c, and d is independently 2-10. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, or 7.

[0413] In some embodiments, a is 2, b is 2, c is 4, and d is 7.

[0414] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0415] In some embodiments, an oligonucleotide comprises mN fN mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide is mN fN mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0416] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0417] In some embodiments, an oligonucleotide comprises mN * fN * mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * fN * mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0418] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN mN * mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotideAttorney Docket No.: 2010794-3211comprises an antisense strand that is mN * fN * mN mN igN fN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0419] In some embodiments, a pattern of sugar modifications comprises mN fN (mN), igN mN (fN)b (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN), igN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0420] In some embodiments, an oligonucleotide comprises mN fN (mN), igN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide is mN fN (mN), igN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand comprises mN fN (mN), igN mN (fN)b (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an antisense strand is mN fN (mN)aigN mN (fN)b (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0421] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN (mN)aigN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN (mN), igN mN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-OMe modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0422] In some embodiments, each of a, b, c, and d is as described herein.

[0423] In some embodiments, a sum of a, b, c, and d is not greater than 30. In some embodiments, a sum of a, b, c, and d is not greater than 25. In some embodiments, a sum of a, b, c, and d is not greater than 24. In some embodiments, a sum of a, b, c, and d is not greater than 23. In some embodiments, a sum of a, b, c, and d is not greater than 22. In some embodiments, a sum of a, b, c, and d is not greater than 21. InAttorney Docket No.: 2010794-3211some embodiments, a sum of a, b, c, and d is not greater than 20. In some embodiments, a sum of a, b, c, and d is not greater than 19. In some embodiments, a sum of a, b, c, and d is not greater than 18. In some embodiments, a sum of a, b, c, and d is not greater than 17. In some embodiments, a sum of a, b, c, and d is not greater than 16. In some embodiments, a sum of a, b, c, and d is not greater than 15. In some embodiments, a sum of a, b, c, and d is not greater than 14. In some embodiments, a sum of a, b, c, and d is not greater than 13. In some embodiments, a sum of a, b, c, and d is not greater than 12. In some embodiments, a sum of a, b, c, and d is not greater than 11. In some embodiments, a sum of a, b, c, and d is not greater than 10.

[0424] In some embodiments, each of a, b, c, and d is independently 1-10. In some embodiments, each of a, b, c, and d is independently 2-10. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, 7, or 8. In some embodiments, each of a, b, c, and d is independently an integer selected from 2, 3, 4, 5, 6, or 7.

[0425] In some embodiments, a is 3, b is 2, c is 4, and d is 7.

[0426] In some embodiments, a pattern of sugar modifications comprises mN fN mN mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, a pattern of sugar modifications is mN fN mN mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0427] In some embodiments, an oligonucleotide comprises mN fN mN mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide is mN fN mN mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an antisense strand comprises mN fN mN mN mN igN mN fN fN mN mN mN mN fN mN f mN mN mN mN mN mN mN. In some embodiments, an antisense strand is mN fN mN mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0428] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN fN mN mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN. In some embodiments, an oligonucleotide comprises an antisense strand that is mN fN mN mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN mN mN.

[0429] In some embodiments, an oligonucleotide comprises mN * fN * mN mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide is mN * fN * mN mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand comprises mN * fN * mN mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an antisense strand is mN * fN * mNAttorney Docket No.: 2010794-3211mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0430] In some embodiments, an oligonucleotide comprises an antisense strand comprising mN * fN * mN mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage. In some embodiments, an oligonucleotide comprises an antisense strand that is mN * fN * mN mN mN igN mN fN fN mN mN mN mN fN mN fN mN mN mN mN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

[0431] In some embodiments, a pattern of sugar modifications comprises mN fN (mN)afN igN (fN)b (mN)c fN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodiments, a pattern of sugar modifications is mN fN (mN), fN igN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater.

[0432] In some embodiments, an oligonucleotide comprises mN fN (mN)afN igN (fN)b (mN)cfN mN fN (mN)d, wherein N represents any nucleoside, m represents a 2’-0Me modification, f represents a 2’-F modification, igN represents a GNA-iso nucleoside, and each of a, b, c, and d are independently an integer of 1 or greater. In some embodimen...

Claims

Attorney Docket No.: 2010794-3211CLAIMS1. An oligonucleotide comprising a sense strand and an antisense strand, wherein:the antisense strand comprises mN fN (mN), fN mN (fN)b (mN), fN mN fN (mN)d fN (mN)e, wherein:m represents a 2’-0Me modification,f represents a 2’-F modification,N represents any nucleoside,a is an integer of 1 or greater,b, c, d, and e are each independently an integer of 2 or greater, andthe sum of a, b, c, d. and e is not greater than 22;or a pharmaceutically acceptable salt thereof.

2. An oligonucleotide comprising a sense strand and an antisense strand, wherein:the sense strand comprises fN mN fN (mN)gfN mN (fN)h (mN); fN (mN)j, wherein:m represents a 2’-0Me modification,f represents a 2’-F modification,N represents any nucleoside,g, h, i, and j are each independently an integer of 2 or greater, andthe sum of g, h, i, and j is not greater than 24;or a pharmaceutically acceptable salt thereof.

3. An oligonucleotide comprising a sense strand and an antisense strand, wherein:the antisense strand comprises mN fN (mN), fN mN (fN)b(mN)cfN mN fN (mN)a IN (mN)eand the sense strand comprises fN mN fN (mN)gfN mN (fN)h (mN); fN (mN)j, wherein:m represents a 2’-0Me modification,f represents a 2’-F modification,N represents any nucleoside,a is an integer of 1 or greater,b, c, d, e, g, h, i, and j are each independently an integer of 2 or greater,the sum of a, b, c, d, and e is not greater than 22, andthe sum of g, h, i, and j is not greater than 24;or a pharmaceutically acceptable salt thereof.Attorney Docket No.: 2010794-32114. The oligonucleotide of any one of the preceding claims, wherein the antisense strand comprises mN fN mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN mN mN.

5. The oligonucleotide of any one of the preceding claims, wherein the sense strand comprises fN mN fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN mN mN.

6. The oligonucleotide of any one of the preceding claims, wherein the antisense strand comprises mN * fN * mN mN mN fN mN fN fN mN mN mN mN fN mN fN mN mN mN fN mN * mN * mN, wherein * represents a phosphorothioate internucleotidic linkage.

7. The oligonucleotide of any one of the preceding claims, wherein the sense strand comprises fN * mN * fN mN mN mN fN mN fN fN fN mN mN mN fN mN mN mN mN * mN * mN.

8. The oligonucleotide of any one of the preceding claims, wherein the oligonucleotide is a pharmaceutically acceptable salt.

9. A composition comprising an oligonucleotide of any one of the preceding claims.

10. A pharmaceutical composition comprising an oligonucleotide of any one of claims 1-8 and a pharmaceutically acceptable carrier.

11. A method, comprising administering or delivering to a system an oligonucleotide of any one of claims 1-8, a composition of claim 9, or a pharmaceutical composition of claim 10.

12. A method for reducing a level of a target RNA, a level of a polypeptide encoded by a target RNA, and / or a level of activity of a protein in a system, comprising administering or delivering to the system an oligonucleotide of any one of claims 1-8, a composition of claim 9, or a pharmaceutical composition of claim 10.

13. A method for treating or preventing a condition, disorder, or disease, comprising administering or delivering to a subject suffering therefrom or susceptible thereto a therapeutically effective amount of an oligonucleotide of any one of claims 1-8, a composition of claim 9, or a pharmaceutical composition of claim 10.Attorney Docket No.: 2010794-321114. Use of an oligonucleotide of any one of claims 1-8, a composition of claim 9, or a pharmaceutical composition of claim 10 in a method of any one of claims 11-13.

15. Use of an oligonucleotide of any one of claims 1-8, a composition of claim 9, or a pharmaceutical composition of claim 10 in the manufacture of a medicament for treating or preventing a condition, disorder, or disease.

16. An oligonucleotide of any one of claims 1-8, a composition of claim 9, or a pharmaceutical composition of claim 10 for use in a method of any one of claims 11-13.

17. A method for preparing an oligonucleotide of any one of claims 1-8, a composition of claim 9, or a pharmaceutical composition of claim 10, comprising one or more synthetic cycles.

18. An oligonucleotide, composition, pharmaceutical composition, method, or use of any one of Embodiments 1-209.

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