214 results about "Modified nucleosides" patented technology

The technology described herein relates to 5'-modified nucleosides, nucleotides, oligonucleotides and double-stranded RNAs, e.g., siRNAs, and kits comprising them and methods of their use for inhibiting target genes.

Provided is an siRNA for inhibiting the gene expression of complement factor B (CFB), or a pharmaceutical composition or conjugate containing the siRNA. The siRNA contains a sense strand and an antisense strand. Each nucleotide in the siRNA is an independent modified or unmodified nucleotide. The sense strand contains a segment of nucleotide sequence I, and the nucleotide sequence I and a nucleotide sequence show in SEQ ID NO: 1 have an equal length and differ by no more than 3 bases. The antisense strand contains a nucleotide sequence II, and the nucleotide sequence II and a nucleotide sequence shown in SEQ ID NO: 2 have an equal length and differ by no more than 3 bases. The siRNA or the pharmaceutical composition or conjugate containing the siRNA can effectively treat and / or prevent diseases related to the mRNA level of the gene expression of the CFB.

The present disclosure provides modified nucleosides, nucleotides, and nucleic acids, and methods of using thereof.

The present invention relates to variants of Terminal deoxynucleotidyl Transferase (TdT), each of which (i) has an amino acid sequence similarity to SEQ ID NO: 2, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33 or 35 with corresponding amino acid substitutions, (ii) is capable of synthesizing a nucleic acid fragment without a template and (iii) is capable of incorporating a modified nucleotide into the nucleic acid fragment.

The invention belongs to the field of biological medicine, and relates to siRNA and a conjugate for inhibiting APP gene expression, the siRNA comprises a positive-sense strand and an antisense strand, the positive-sense strand comprises a nucleotide sequence I, and the antisense strand comprises a nucleotide sequence II; each nucleotide in the nucleotide sequence I and the nucleotide sequence II is modified or unmodified nucleotide; the nucleotide sequence I and the nucleotide sequence II are at least partially reversely complementary to form a double-stranded region; the nucleotide sequence I is basically consistent with a first section of nucleotide sequence, and the first section of nucleotide sequence is a section of nucleotide sequence with the length of at least 19 nucleotides in mRNA expressed by an APP gene. The siRNA can specifically induce degradation of APP mRNA, so that synthesis of APP in the liver is inhibited, APP protein is induced to be reduced durably, pathological deposition of Abeta and other related toxic proteins is reduced, and the siRNA has a good patent medicine prospect.

The present disclosure relates to novel destabilizing molecules, such as a butadiol moiety, modified nucleosides incorporating the butadiol moiety, modified nucleotides incorporating the butadiol moiety, and oligonucleotides and RNAi agents comprising such modified nucleotides that improve selectivity and / or reduce off target effects while maintaining or improving knockdown of on target gene expression.

An siRNA capable of inhibiting the expression of apolipoprotein C3 (APOC3) gene, comprising a sense strand and an antisense strand, wherein the sense strand and the antisense strand are respectively composed of nucleotide sequence I or nucleotide sequence II comprising 19 modified or unmodified nucleotides, and the nucleotide sequence I and the nucleotide sequence II are at least partially reverse complementary to form a double-stranded region, and the nucleotide sequence II is at least partially reverse complementary to a nucleotide sequence segment in the mRNA of APOC3 gene expression; in the direction from the 5'-end to the 3'-end, at least one of the 3-6th nucleotides of the nucleotide sequence II is a stabilized modified nucleotide. The siRNA as described above, as well as the pharmaceutical composition and siRNA conjugate comprising the siRNA, can effectively treat and / or prevent diseases or disorders related to APOC3 gene expression, and have a significantly reduced off-target effect.

Described herein are modified RNA molecules where a 3′-stabilizing region is covalently attached to the RNA, and where the 3′-stabilizing region comprises one or more modified nucleosides. Methods of synthesizing said RNAs are also provided herein.

The invention provides a nucleoside phosphoramidite compound as shown in a formula I. The invention further provides a preparation method of the nucleoside phosphoramidite compound. The nucleoside phosphoramidite compound disclosed by the invention can be introduced into the head and tail ends and / or the middle of oligonucleotide, the stability of the oligonucleotide is enhanced while the biological function of the oligonucleotide is enhanced or basically maintained, and the continuous acting time is prolonged.

The present disclosure relates to a nucleoside analogue and a use thereof. The nucleoside analogue is a compound shown in formula (Ia) or a prodrug thereof. The nucleoside analogue can enhance targeted delivery of oligonucleotide drugs to nervous system cells, thereby increasing the inhibition rate of the oligonucleotide drugs delivered in a targeted manner on expression of specific genes in the nervous system cells, or enabling the oligonucleotide drugs delivered in a targeted manner to achieve the purpose of preventing and / or treating pathological conditions or diseases caused by abnormal expression of the specific genes in the nervous system cells. The present disclosure also relates to a nucleotide analogue-containing double-stranded oligonucleotide and a use thereof. The double-stranded oligonucleotide comprises a sense strand and an antisense strand, each strand has 17-25 modified and / or unmodified nucleotides, and the antisense strand and the sense strand are complementary to form a duplex region; the sense strand and / or the antisense strand contains at least one nucleotide analogue represented by the structure of formula (100), or a tautomer, or stereoisomer, or pharmaceutically acceptable salt thereof; the double-stranded oligonucleotide can effectively treat and / or prevent pathological conditions or diseases caused by abnormal expression of specific genes in nervous system cells.

The invention discloses base-modified mRNA (messenger Ribonucleic Acid) and application thereof in a cell-free protein synthesis system, and belongs to the technical field of nucleic acid modification. Natural nucleoside triphosphate is completely replaced by different types of basic group modified nucleoside triphosphate, mRNA with different chemical modifications is obtained through an in-vitro transcription method and purification, and the mRNA is put into an in-vitro cell-free protein synthesis system for reaction. A reaction element recombination system-PURE system based on prokaryote Escherichia coli purified protein and an eukaryote CFPS system taking rabbit reticulated red lysate as a material are mainly selected to detect the corresponding translation efficiency of mRNA modified by different bases in two cell-free protein synthesis systems respectively. Different regulatory activities may be shown in different cell-free protein synthesis systems when different modified nucleotides are doped into mRNA, base modified triphosphoric acid capable of improving the translation efficiency of an in-vitro protein synthesis system is screened out, and the stability of mRNA in the cell-free protein synthesis system and the corresponding protein expression level are improved.

Provided are a double‑stranded RNA composition for modulating DGAT2 expression, a conjugate thereof, a pharmaceutical composition comprising same, and a use thereof. The double-stranded RNA comprises a sense strand and an antisense strand, and each nucleotide in the double-stranded RNA is independently a modified or unmodified nucleotide; the sense strand comprises one nucleotide sequence selected from the nucleotide sequences shown in SEQ ID NOs: 1-86 or a nucleotide sequence having no more than 3 nucleotide differences from the above sequences, and the antisense strand comprises one nucleotide sequence selected from the nucleotide sequences shown in SEQ ID NOs: 87-172 or a nucleotide sequence having no more than 5 nucleotide differences from the above sequences. The double-stranded RNA, the conjugate thereof and the pharmaceutical composition comprising same can be delivered to the liver by means of small nucleic acid delivery technology, and are expected to provide a safer and more effective therapeutic plan for patients with MASH-associated metabolic diseases.

This disclosure relates to the modified 3' region elements that confer improved therapeutic properties including, but not limited to, increased stability, increased expression, and / or a reduced innate immune response when introduced into a population of cells, even in the absence of a 5'-cap. The disclosure also provides polynucleotides (e.g., RNA molecules) containing modified 3' regions. In some embodiments of the disclosure, the modified 3' regions contain modified adenosines (e.g., 2'-0-methoxy- adenosine, 2'-fluoro-adenosine, or locked nucleic acid (LNA)-adenosine) and / or a modified thymidine (e.g., inverted deoxythymidine). In some embodiments of the disclosure, the modified 3' region may contain one or more modified internucleoside linkages (e.g., a phosphorothioate linkage) or modified nucleobase (e.g., N1-methyl-pseudouridine). The polynucleotides of the disclosure may be used to express a desired polypeptide in a subject, such as a subject having a disease or condition associated with a deficiency in the corresponding endogenous polypeptide.

The disclosure relates to a method for sequencing of a DNA strand, the method includes (S200) providing a double-stranded DNA fragment with a first strand and a second strand, the DNA strand to be sequenced corresponding to the first strand or the second strand, (S300A) connecting the first and second strands with a first hairpin oligonucleotide and a second hairpin oligonucleotide, (S500A) amplifying the ligated product obtained in step (S300A) using rolling circle amplification (RCA), (S600A) amplifying the ligated product obtained in step (S300A) using RCA or the amplicons obtained in step (S500A) using primer extension using a first group of redox-modified nucleotides and a second group of redox-modified nucleotides, and (S800A) determining the sequence of the DNA strand by sequencing the amplicons (80) obtained in step (S600A) using an electrochemical sequencing method.

Provided herein are RNA molecules (e.g. self-amplifying messenger ribonucleic acids (samRNA)) comprising modified nucleotides, such as N1-methyl-pseudouridine.

This application relates to mutant polymerases. This application discloses mutant phage-type RNA polymerases, such as a mutant T7, SP6, and T3 RNA polymerase, may use 2′-modified nucleoside triphosphates or deoxynucleotide triphosphates as substrates. Methods for producing nucleic acid molecules using these mutant polymerases are also disclosed.

A single-stranded oligonucleotide having a length of 16-30 nucleotides. The single-stranded oligonucleotide and complement component C3 (CC3) mRNA have sufficient complementarity to mediate RNAi effect; each nucleotide in the single-stranded oligonucleotide is a modified or unmodified nucleotide, at least one nucleotide in the single-stranded oligonucleotide is a nucleotide X, and the at least one nucleotide is a fluoro-modified nucleotide; moreover, according to a direction from the 5' end to the 3' end, at least one nucleotide X is located after an eighth nucleotide and is spaced apart from the eighth nucleotide in the single-stranded oligonucleotide by 4-7 nucleotides; and each nucleotide X is a deoxynucleotide or an unmodified nucleotide. The present invention further relates to a double-stranded oligonucleotide comprising the single-stranded oligonucleotide as an antisense strand, an oligonucleotide conjugate, and a pharmaceutical composition.

A single-stranded oligonucleotide having a length of 16-30 nucleotides. The composition of the single-stranded oligonucleotide enables the single-stranded oligonucleotide to inhibit the expression of target mRNA by means of an RNAi mechanism. Each nucleotide in the single-stranded oligonucleotide is independently a modified or unmodified nucleotide, wherein at least one nucleotide in the single-stranded oligonucleotide is a nucleotide X; at least one nucleotide is a fluoro-modified nucleotide; and, in the 5' to 3' direction, the 13th nucleotide of the single-stranded oligonucleotide is a substituted alkoxy-modified nucleotide, the 14th nucleotide of the single-stranded oligonucleotide is a nucleotide X, and each of the 15th nucleotide and all subsequent nucleotides of the single-stranded oligonucleotide is independently a modified nucleotide. A double-stranded oligonucleotide comprising the single-stranded oligonucleotide as an antisense strand, an oligonucleotide conjugate and a pharmaceutical composition.

Disclosed herein are improved methods and compositions of de novo synthesis of nucleic acids by cyclic extension using nucleotides that have an N-linked or O-linked scar or protecting group, and subsequent removal of the scar or protecting group. During polymerase-nucleotide conjugate-based polynucleotide synthesis or after the synthesis is completed, nucleobases having a scar from linker cleavage can be converted back into native form by removal of the scars from the nucleobases. Secondary structure formation in the nascent chain, which may inhibit extension reactions, is suppressed by the use of monomers with protected nucleobases that prevent Watson-Crick base pairing and / or other structures. After the synthesis is completed, the nucleobases can be converted back into native form by palladium-based removal of the protecting groups.

The present invention relates to the detection of modifications in polynucleotides by binding an affinity binding element to a nucleotide sequence. The affinity binding element may include a compound that binds to a modified nucleotide base and a nucleotide sequence complementary to a target nucleotide sequence, an oligonucleotide primer complementary to the target nucleotide sequence, or a nucleotide sequence complementary to an oligonucleotide linker. The present invention provides methods for detecting one or more modifications in a nucleotide sequence. The invention also provides affinity binding elements, and optionally, oligonucleotide primers and / or oligonucleotide probes, and methods of using the affinity binding elements in assays to detect modified bases. The methods of the invention enable the detection of modified nucleotides associated with cancer, psychiatric disorders or metabolic diseases.

The present disclosure relates to a 5'-modified nucleoside derivative and an oligonucleotide comprising same. Specifically, the oligonucleotide comprises at least one structure represented by formula (I), wherein each functional group is defined in the context of the present disclosure.

To provide compounds, methods, and pharmaceutical compositions for reducing the amount or activity of leucine-rich repeat kinase (LRRK2) RNA in a cell or animal.SOLUTION: There is provided an oligomeric compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides. Therein: a nucleobase sequence of the modified oligonucleotide is at least 90% complementary to an equal length portion of a LRRK2 nucleic acid; and the modified oligonucleotide comprises at least one modification selected from a modified sugar, a sugar surrogate, and a modified internucleoside linkage.SELECTED DRAWING: None

Provided herein is an antisense oligonucleotide for targeting long non-coding RNA maternally expressed gene 3 (IncRNA Meg3), wherein the oligonucleotide comprises or consists of one of the following sequences in the 5' to 3' direction: (I) C G C dC dG dC dC dA dT dA dT dC dT C C C (SEQ I D NO: 1 ); (ii) C G G dA dC dA dA dA dA dC dT dG dG T T G (SEQ ID NO: 2); (ill) A A G dA dG dT dC dT dC dC dT dC dC dT T A A (SEQ ID NO: 3); (iv) G A T dT dA dG dC dC dC dT dG dT dG dT T C A (SEQ ID NO: 4); (v) C G G dC dA dC dA dA dG dA dG dC dC dA A A G (SEQ ID NO: 5); (vi) T O T dC dC dT dC dC dT dT dA dA dG dC C C C (SEQ ID NO: 6); or (vii) G T C dT dC dC dT dC dC dT dT dA dA dG C C C (SEQ ID NO: 7); wherein dA, dT, dG and dC are deoxyribonucleotides, or wherein dC is 5-methyl-2'-deoxycytidine; and wherein A, T, G and C are modified or unmodified nucleotide building blocks.

To provide a closed linear DNA containing modified nucleotides which is particularly useful for a therapeutic purpose.SOLUTION: Provided is a closed linear DNA (clDNA) comprising stem regions including a target double-stranded DNA sequence which is covalently closed at both ends by a hairpin loop, and includes at least two modified nucleotides.SELECTED DRAWING: Figure 1

Disclosed herein are modified RNAs comprising one or more one or more modified nucleotides at position +1 to position +6 with reference to a 5' terminus of the RNA, and methods of making the same. Also provided are compositions comprising one or more of the modified RNAs provided herein, and methods of using said compositions for therapeutic applications.

The invention discloses a preparation method of a 2 '-modified ribose derivative, 2'-modified nucleoside is taken as a raw material, hydroxyl is subjected to benzoyl protection, basic groups are removed by a one-step method, and 1-site is acetylated, so that the ribose derivative with wide application is obtained, the preparation method is simple, the reaction condition is mild, the synthesis efficiency is high, and the preparation method is environment-friendly. The method is suitable for large-scale production, and the preparation method has good substrate applicability.

The invention relates to a small interfering nucleic acid for inhibiting FGFR1 (Fibroblast Growth Factor Receptor 1) gene expression and pharmaceutical application thereof. The small interfering nucleic acid comprises a positive-sense strand and an antisense strand, all nucleotides in the positive-sense strand and the antisense strand are modified nucleotides, and the positive-sense strand and the antisense strand are reversely complementary to form a double-strand region. The small interfering nucleic acid can be used for preparing medicines for preventing and treating diseases related to FGFR1 gene expression, and the diseases are selected from bladder cancer, liver tumor, colon cancer, lung cancer, cervical cancer and the like.

Compounds comprising 5-position modified pyrimidines are provided. Further, polynucleotides, such as aptamers, comprising at least one 5-position modified pyrimidine are provided. Methods of selecting and using such polynucleotides, such as aptamers, are also provided.