Gene editing methods and compositions for eliminating risk of jc virus activation and pml (progressive multifocal leukoencephalopathy) during immunosuppressive therapy

Abstract

A method of eliminating the risk of JCV activation in a subject undergoing immunosuppressive therapy, by administering an effective amount of a gene editing composition directed toward at least one target sequence in the JCV genome, cleaving the target sequence in the JCV genome, disrupting the JCV genome, eliminating the JCV infection, eliminating the risk of JCV activation, and treating the subject with an immunosuppressive therapy. A pharmaceutical composition including at least one isolated nucleic acid sequence encoding a CRISPR-associated endonuclease and at least one gRNA having a spacer sequence complementary to a target sequence in a JCV DNA, the isolated nucleic acid sequences being included in at least one expression vector. Pharmaceutical compositions including at least one isolated nucleic acid sequence encoding at least one TALEN, at least one ZFN, and gene editing composition of C2c1, C2c3, TevCas9, Archaea Cas9, CasY.1-CasY.6, CasX, or argonaute protein, which target at least one nucleotide sequence of the JCV genome.

1 - 23 . (canceled) 
     
     
         24 . A method of inactivating JC virus (JCV) in a cell comprising: the method comprising:
 delivering to the cell a nucleic acid encoding:   (a) a CRISPR-associated endonuclease Cas9 or a nucleic acid sequence encoding the CRISPR-associated endonuclease Cas9; and   (b) a guide RNA (gRNA) or a nucleic acid sequence encoding the first gRNA, the guide RNA being complementary to a target sequence within the JCV VP1 gene.   
     
     
         25 . The method of  claim 24 , wherein the CRISPR-associated endonuclease Cas9 is selected from the group consisting of a wild-type Cas9, a human-optimized Cas9, and a nickase mutant Cas9. 
     
     
         26 . The pharmaceutical composition of  claim 25 , wherein the CRISPR-associated endonuclease Cas 9 is a  Staphylococcus aureus  Cas9 (SaCas9). 
     
     
         27 . A nucleic acid molecule encoding:
 (a) a CRISPR-associated endonuclease Cas9 or a nucleic acid sequence encoding the CRISPR-associated endonuclease Cas9; and   (b) a guide RNA (gRNA) or a nucleic acid sequence encoding the first gRNA, the guide RNA being complementary to a target sequence within the JCV VP1 gene.   
     
     
         28 . The nucleic acid molecule of  claim 27 , wherein the CRISPR-associated endonuclease Cas9 is selected from the group consisting of a wild-type Cas9, a human-optimized Cas9, and a nickase mutant Cas9. 
     
     
         29 . The pharmaceutical composition of  claim 28 , wherein the CRISPR-associated endonuclease Cas9 is a  Staphylococcus aureus  Cas9 (SaCas9).