Rna guided eradication of herpes simplex type i and other related herpesviruses
AssigneeTemple University of the Commonwealth System of Higher Education
InventorsPamela C. Roehm, Kamel Khalili, Masoud Sayed Shekarabi, Hassen Wollebo
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Abstract
The present invention relates to compositions and methods for the inhibition of the infectivity of a herpesvirus. The compositions and methods provide a CRISPR-associated (Cas) peptide and a guide nucleic acid, which induce the mutation of herpesvirus genome, thereby inhibiting the infectivity of the herpesvirus. Further disclosed are Cas peptides including Cas9 or a variant thereof comprising one or more point mutations relative to wildtype Streptococcus pyogenes Cas9 (spCas9), and Cpf1 or a variant thereof.
Claims20
An expression vector comprising a nucleic acid sequence encoding:
- (a) a Cas protein;
- and
- (b) a first guide RNA (gRNA) and a second gRNA molecule,
- each gRNA comprising a nucleotide sequence substantially complementary to a different target sequence in a herpesvirus genome, the first gRNA comprising a spacer sequence of any one of SEQ ID NOs:
- 34, 35, or 36 or a fragment thereof and being substantially complementary to a first target sequence within an ICP0 region of the herpesvirus genome, and the second gRNA being substantially complementary to a second target sequence within an ICP4 or an ICP27 region of the herpesvirus genome.
A pharmaceutical composition comprising:
- an expression vector comprising a nucleic acid sequence encoding:
- (a) a Cas protein;
- and
- (b) first guide RNA (gRNA) and a second gRNA molecule,
- each gRNA comprising a nucleotide sequence substantially complementary to a different target sequence in a herpesvirus genome, the first gRNA comprising a spacer sequence of any one of SEQ ID NOs:
- 34, 35, or 36 or a fragment thereof and being substantially complementary to a first target sequence within an ICP0 region of the herpesvirus genome, and the second gRNA being substantially complementary to a second target sequence within an ICP4 or an ICP27 region of the herpesvirus genome.
A method of inactivating a viral deoxyribonucleic acid (DNA) sequence of a herpesvirus genome in a cell, the method comprising contacting the cell with a therapeutically effective amount of a composition comprising:
- (a) a Cas protein or a nucleic acid encoding a Cas protein;
- and
- (b) first guide RNA (gRNA) and a second gRNA molecule,
- each gRNA comprising a nucleotide sequence substantially complementary to a different target sequence in the herpesvirus genome, the first gRNA comprising a spacer sequence of any one of SEQ ID NOs:
- 34, 35, or 36 or a fragment thereof and being substantially complementary to a first target sequence within an ICP0 region of the herpesvirus genome, and the second gRNA being substantially complementary to a second target sequence within an ICP4 or an ICP27 region of the herpesvirus genome.
Dependencies are inferred from the claim text.