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Kafe 바로가기국가/구분 | United States(US) Patent 등록 |
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국제특허분류(IPC7판) |
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출원번호 | US-0143364 (2016-04-29) |
등록번호 | US-9657295 (2017-05-23) |
발명자 / 주소 |
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출원인 / 주소 |
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대리인 / 주소 |
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인용정보 | 피인용 횟수 : 2 인용 특허 : 381 |
The present disclosure provides modified nucleosides, nucleotides, and nucleic acids, and methods of using thereof.
1. A method of synthesizing a messenger ribonucleic acid (mRNA), comprising the steps of: a) providing a complementary deoxyribonucleic acid (cDNA) that encodes a pharmaceutical protein of interest;b) selecting a nucleotide that disrupts a binding of a major groove binding partner with the RNA, wher
1. A method of synthesizing a messenger ribonucleic acid (mRNA), comprising the steps of: a) providing a complementary deoxyribonucleic acid (cDNA) that encodes a pharmaceutical protein of interest;b) selecting a nucleotide that disrupts a binding of a major groove binding partner with the RNA, wherein the nucleotide has decreased binding affinity to the major groove binding partner selected from the group consisting of toll-like receptor (TLR) 3, TLR7, TLR8, retinoic acid-inducible gene I (RIG-I), melanoma differentiation-associated gene 5 (MDA5) and laboratory of genetics and physiology 2 (LGP2), and wherein the nucleotide comprises a modification on the major groove face of the nucleobase where an atom of the major groove face of the nucleobase is replaced or substituted with an alkyl group;c) contacting the provided cDNA and the selected nucleotide with an RNA polymerase under conditions such that an RNA transcript is synthesized, andd) 5′-capping the RNA transcript concomitantly or post-transcriptionally such that the mRNA is synthesized. 2. The method of claim 1, wherein the 5′-capping is performed concomitantly. 3. The method of claim 1, wherein the nucleobase comprising a modification on the major groove face is a pyrimidine nucleobase. 4. The method of claim 3, wherein the pyrimidine nucleobase is selected from cytosine and uracil. 5. The method of claim 3, wherein the pyrimidine nucleobase is uracil. 6. The method of claim 3, wherein the pyrimidine nucleobase is cytosine. 7. The method of claim 5, wherein the nucleotide comprises 1-methyl-pseudouridine or 5-methyl-uridine. 8. The method of claim 7, wherein the nucleotide comprises 1-methyl-pseudouridine. 9. The method of claim 6, wherein the nucleotide comprises 5-methyl-cytidine. 10. The method of claim 1, wherein the major groove binding partner is TLR3, TLR7, or TLR8. 11. The method of claim 1, wherein the major groove binding partner is RIG-I, MDA5, or LGP2. 12. The method of claim 2, wherein the nucleobase comprising a modification on the major groove face is a pyrimidine nucleobase. 13. The method of claim 12, wherein the pyrimidine nucleobase is selected from cytosine and uracil. 14. The method of claim 12, wherein the pyrimidine nucleobase is uracil. 15. The method of claim 12, wherein the pyrimidine nucleobase is cytosine. 16. The method of claim 14, wherein the nucleotide comprises 1-methyl-pseudouridine or 5-methyl-uridine. 17. The method of claim 16, wherein the nucleotide comprises 1-methyl-pseudouridine. 18. The method of claim 15, wherein the nucleotide comprises 5-methyl-cytidine. 19. The method of claim 2, wherein the major groove binding partner is TLR3, TLR7, or TLR8. 20. The method of claim 2, wherein the major groove binding partner is RIG-I, MDA5, or LGP2.
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