US 11,643,672 B2 · Inducible CRISPR system expression and applications thereof

Abstract

The invention pertains to an inducible CRISPR system for controlling expression of a CRISPR complex with an inducible fusion promoter. One embodiment of the invention provides HIV LTR-minimal Drosophila hsp70 fusion promoter that can be used for inducible co-expression of gRNA and Cas9 in HIV-infected cells to target cellular cofactors such as Cyclin T1. A single introduction of such embodiment leads to sustained suppression of HIV replication in stringent, chronically infected HeLa-CD4 cell lines as well as in T-cell lines. In another embodiment, the invention further relates to enhancement of HIV suppression by incorporating cis-acting ribozymes immediately upstream of the gRNA in the inducible CRISPR system construct. The inducible fusion promoter is adaptable for other tissue- or cell-type specific expression of the inducible CRISPR system.

Claims15

Dependent claims

An inducible Clustered, Regularly Interspaced, Short Palindromic Repeats (CRISPR) system comprising:

an inducible fusion promoter comprising a minimal heat shock promoter that is operatively associated with an inducible element;
a nucleotide sequence encoding a guide sequence;
and
a nucleotide sequence encoding a CRISPR-associated protein (Cas),
said inducible fusion promoter being inducible by an inducer to drive expression of said guide sequence and said Cas, and
said inducible CRISPR system comprising a nucleotide sequence encoding a modified ribozyme having lower cleavage efficiency than before the modification, said nucleotide sequence encoding a modified ribozyme being SEQ ID NO:
16 or SEQ ID NO:
17.

5 more elements

Dependencies are inferred from the claim text.

Inducible CRISPR system expression and applications thereof

Life of this patent

Inducible CRISPR system expression and applications thereof

Life of this patent

Abstract

Claims15