163 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.

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.

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.

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.

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.

Disclosed are a modified cytidine compound, i.e. a new derivative cytidine generated by adding a guanidyl at position 4 of a cytidine, and a nucleic acid containing the modified compound, for example, RNA. The nucleic acid containing the modified cytidine, especially mRNA, can significantly increase the expression quantity of the mRNA in vivo.

The present disclosure provides novel self-amplifying RNA (saRNA) constructs that demonstrate enhanced protein expression, prolonged durability, reduced immunogenicity, and the ability to express multiple therapeutic proteins homogeneously. The saRNA constructs comprise a 5' untranslated region (5'UTR), non-structural protein genes derived from alphaviruses, at least one gene of interest encoding a therapeutic protein, a 3' untranslated region (3'UTR), and one or more modified nucleosides. Also disclosed are dual construct systems comprising a first construct encoding non-structural proteins and a second construct encoding one or more genes of interest. Methods of producing and using the saRNA constructs for engineering cells, particularly immune cells, for treatment of various conditions including cancer, inflammatory conditions, and infectious diseases are provided. The saRNA constructs enable the generation of "armored" immune cells expressing multiple therapeutic proteins, thereby providing a multi-pronged approach to complex diseases.

Methods for detecting modified nucleobases in a DNA sample by excising a modified nucleobase of interest using a specific DNA glycosylase are described. Prior to the glycosylase treatment, the DNA target fragment is copied by a DNA polymerase to produce a complementary copy strand that retains genetic information of the DNA target strand. After glycosylase treatment, the DNA target fragment is repaired by: linking across a nick to produce a deletion at each position of the modified nucleobase of interest; or filling the nicks with a single non-natural nucleotide to produce a base substitution at each position of the modified nucleobase of interest. The comparison of the DNA sequences of the two strands of the target fragment enables the recognition of the positions of the modified nucleotide bases in the DNA target fragment.

A nucleoside phosphoramidite compound, the compound being a 2'-phosphoramidite group, 4'-modified TNA structured compound. The nucleoside phosphoramidite compound can be used to incorporate into the first and / or last and / or intermediate of an oligonucleotide such that the modified oligonucleotide hybridizes to a portion of a target mRNA, thereby causing a loss or down-regulation of normal function of the target mRNA.

Described are methods of detecting modified nucleotide bases in a DNA sample using specific DNA glycosylases to excise a modified nucleobase of interest. Prior to glycosylase treatment, DNA target fragments are copied by a DNA polymerase to produce a complementary copy strand that preserves the genetic information of the DNA target strand. Following glycosylase treatment, the DNA target fragments are repaired by either ligating across the gaps to produce a deletion at each position of the modified nucleobase of interest or filling in the gaps with a single non-native nucleotide to produce a base substitution at each position of the modified nucleobase of interest. Comparison of the DNA sequences of the two strands of the target fragments enables identification of the positions of the modified nucleotide base in the DNA target fragment.

This invention provides the application of a class of nucleoside compounds that effectively inhibit CCHFV replication, particularly the application of 2'-α-fluoro-2'-β-methyl modified nucleoside compounds in the preparation of drugs for treating CCHF disease. Studies have found that 2'-α-fluoro-2'-β-methyl-CTP can specifically and efficiently inhibit CCHFV L protein polymerase through an immediate chain termination mechanism, but it cannot be incorporated into the L protein polymerases of Lassa virus (LASV) and Rift Valley fever virus (RVFV), which belong to the same order Arenavirales but different families as CCHFV. Further introduction of this modification into ATP, UTP, and GTP also exhibits good immediate chain termination activity. This invention provides a new direction for the preparation of anti-CCHFV nucleoside drugs and demonstrates its feasibility.

The present invention relates to 5'-modified nucleoside analogs and oligonucleotides produced therefrom, and more specifically, to modified nucleosides and their analogs that can be used to be incorporated into the oligonucleotide terminus, which can be bound to double-stranded oligonucleotides (short interfering RNA) or single-stranded oligonucleotides (e.g., antisense oligonucleotides). The oligonucleotides provided herein are expected to result in the loss of normal function of the target RNA by hybridizing to a portion of the target RNA.