Compositions and methods for the targeting of PCSK9
AssigneeScribe Therapeutics Inc.
InventorsJason Fernandes, Sean Higgins, Sarah Denny, Ross White, Emeric Jean Marius Charles, Addison Wright, Benjamin Demaree, Benjamin Oakes
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Abstract
Provided herein are gene repressor systems comprising fusion proteins, such as fusion proteins comprising a DNA binding domain such as a TALE, zinc finger or catalytically-dead CRISPR protein and guide nucleic acid (gRNA), which are useful in the repression of a proprotein convertase subtilisin/kexin Type 9 (PCSK9) gene. Also provided are methods of using such systems to repress transcription of PCSK9.
Claims22
A system comprising a repressor fusion protein and a guide ribonucleic acid (gRNA), wherein the repressor fusion protein comprises:
- (a) a catalytically-dead CasX protein (dCasX);
- (b) a repressor domain (RD1) comprising the sequence of SEQ ID NO:
- 130, 131, 135, or 143, or a sequence having at least 95% identity to the sequence of SEQ ID NO:
- 130, 131, 135, or 143;
- (c) a DNMT3A catalytic domain (DNMT3A);
- and
- (d) a DNMT3L domain (DNMT3L),
- wherein the gRNA comprises a targeting sequence complementary to a proprotein convertase subtilisin/kexin Type 9 (PCSK9) gene target nucleic acid sequence,
- wherein the targeting sequence comprises the sequence of SEQ ID NO:
- 1844, 1852, 1853, 1855, 1858, 1859, 1867, 1869, or 1870,
- wherein the repressor fusion protein is capable of forming a ribonucleoprotein (RNP) with the gRNA;
- and
- wherein the RNP is capable of repressing transcription of the PCSK9 gene upon binding to the PCSK9 gene target nucleic acid sequence.
A composition comprising a first nucleic acid and a second nucleic acid, wherein
- (a) the first nucleic acid comprises a guide ribonucleic acid (gRNA) comprising a targeting sequence complementary to a proprotein convertase subtilisin/kexin Type 9 (PCSK9) gene target nucleic acid sequence, wherein the targeting sequence comprises the sequence of SEQ ID NO:
- 1844, 1852, 1853, 1855, 1858, 1859, 1867, 1869, or 1870, and wherein the gRNA is capable of forming a ribonucleoprotein (RNP) with a repressor fusion protein;
- and
- (b) the second nucleic acid encodes a repressor fusion protein comprising:
- i) a catalytically-dead CasX protein (dCasX);
- ii) a repressor domain (RD1) comprising the sequence of SEQ ID NO:
- 130, 131, 135, or 143, or a sequence having at least 95% identity to the sequence of SEQ ID NO:
- 130, 131, 135, or 143;
- iii) a DNMT3A catalytic domain (DNMT3A);
- and
- iv) a DNMT3L domain (DNMT3L).
A method of treating a PCSK9-related disease or disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective dose of a composition comprising a first nucleic acid and a second nucleic acid, wherein:
- (a) the first nucleic acid comprises a guide ribonucleic acid (gRNA) comprising a targeting sequence complementary to a proprotein convertase subtilisin/kexin Type 9 (PCSK9) gene target nucleic acid sequence, wherein the targeting sequence comprises the sequence of SEQ ID NO:
- 1844, 1852, 1853, 1855, 1858, 1859, 1867, 1869, or 1870, and wherein the gRNA is capable of forming a ribonucleoprotein (RNP) with a repressor fusion protein;
- and
- (b) the second nucleic acid comprises an mRNA encoding a repressor fusion protein comprising:
- i) a catalytically-dead CasX protein (dCasX):
- ii) a repressor domain (RD1) comprising the sequence of SEQ ID NO:
- 130, 131, 135, or 143, or a sequence having at least 95% identity to the sequence of SEQ ID NO:
- 130, 131, 135, or 143;
- iii) a DNMT3A catalytic domain (DNMT3A);
- and
- iv) a DNMT3L domain (DNMT3L).
Dependencies are inferred from the claim text.