Excision of retroviral nucleic acid sequences

Abstract

Compositions for the in vivo delivery of a gene editing CRISPR/Cas9 complex was developed to eliminate integrated retroviral DNA sequences from latently infected human cells and animal disease models

1 .- 46 . (canceled) 
     
     
         47 . An adeno-associated viral vector serotype 9 (AAV9) expression vector, comprising a nucleic acid sequence encoding:
 (a) a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease;   (b) a first guide RNA (gRNA), the first gRNA being complementary to a target sequence within a long terminal repeat (LTR) of a human immunodeficiency virus (HIV) sequence, and wherein the first gRNA comprises a sequence having at least 80% sequence identity to SEQ ID NO: 207; and   (c) a second gRNA, the second gRNA being complementary to a target sequence within a GagD region of the HIV sequence.   
     
     
         48 . The AAV9 expression vector of  claim 47 , wherein the CRISPR-associated endonuclease is Cas9 or a homologue thereof. 
     
     
         49 . The AAV9 expression vector of  claim 47 , wherein the first gRNA comprises a sequence having at least 90% sequence identity to SEQ ID NO: 207. 
     
     
         50 . The AAV9 expression vector of  claim 47 , wherein the first gRNA comprises a sequence according to SEQ ID NO: 207. 
     
     
         51 . An adeno-associated viral vector serotype 9 (AAV9) expression vector, comprising a nucleic acid sequence encoding:
 (a) a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease;   (b) a first guide RNA (gRNA), the first gRNA being complementary to a target sequence within a long terminal repeat (LTR) of a human immunodeficiency virus (HIV) sequence; and   (c) a second gRNA, the second gRNA being complementary to a target sequence within a GagD region of the HIV sequence, and wherein the second gRNA comprises a sequence having at least 80% sequence identity to SEQ ID NO: 92.   
     
     
         52 . The AAV9 expression vector of  claim 51 , wherein the CRISPR-associated endonuclease is Cas9 or a homologue thereof. 
     
     
         53 . The AAV9 expression vector of  claim 51 , wherein the second gRNA comprises a sequence having at least 90% sequence identity to SEQ ID NO: 92. 
     
     
         54 . The AAV9 expression vector of  claim 51 , wherein the second gRNA comprises a sequence according to SEQ ID NO: 92. 
     
     
         55 . An adeno-associated viral vector serotype 9 (AAV9) expression vector, comprising a nucleic acid sequence encoding:
 (a) a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease;   (b) a first guide RNA (gRNA), the first gRNA being complementary to a target sequence within a long terminal repeat (LTR) of a human immunodeficiency virus (HIV) sequence, and wherein the first gRNA comprises a sequence having at least 80% sequence identity to SEQ ID NO: 207; and   (c) a second gRNA, the second gRNA being complementary to a target sequence within a GagD region of the HIV sequence, and wherein the second gRNA comprises a sequence having at least 80% sequence identity SEQ ID NO: 92.   
     
     
         56 . The AAV9 expression vector of  claim 55 , wherein the CRISPR-associated endonuclease is Cas9 or a homologue thereof. 
     
     
         57 . The AAV9 expression vector of  claim 55 , wherein the first gRNA comprises a sequence having at least 90% sequence identity to SEQ ID NO: 207. 
     
     
         58 . The AAV9 expression vector of  claim 55 , wherein the first gRNA comprises a sequence according to SEQ ID NO: 207. 
     
     
         59 . The AAV9 expression vector of  claim 55 , wherein the second gRNA comprises a sequence having at least 90% sequence identity to SEQ ID NO: 92. 
     
     
         60 . The AAV9 expression vector of  claim 55 , wherein the second gRNA comprises a sequence according to SEQ ID NO: 92.