A drug-inducible CRISPR/Cas9 system for gene transcriptional activation

A gene transcription and inducible technology, applied in the field of molecular biology, can solve problems such as the inapplicability of regulatory drugs or methods
CN109207518BActive Publication Date: 2020-12-01INST OF ZOOLOGY CHINESE ACAD OF SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
INST OF ZOOLOGY CHINESE ACAD OF SCI
Publication Date
2020-12-01

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Abstract

The invention relates to the field of molecular biology, in particular to a drug inducible CRISPR / Cas9 system for gene transcription activation. The drug inducible CRISPR / Cas9 system comprises 16-22ntsgRNA targeting a specific gene site and any one of vector / vector combination as (1)-(3) below: (1) a vector expressing fusion protein dCas9-(ERT2)n-ADs or dCas9-ADs-(ERT2)n; (2) a vector combinationexpressing fusion protein dCas9-(NLS) m or dCas9-(ERT2)n and MCP-(ERT2)n-ADs; (iii) a vector combination expressing fusion protein dCas9-(NLS)m-(GCN4)p and scFv-(ERT2)n-ADs. Compared with the prior art, the system of the invention can more effectively realize the regulation of drug-induced gene transcription activation, and has more flexible and convenient operation and lower background activity.It is helpful to control the dynamic biological process effectively and precisely, and has great value for the research and development of biomedicine.
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Description

technical field

[0001] The present invention relates to the field of molecular biology, in particular to a drug-inducible CRISPR / Cas9 system for gene transcription activation. Background technique

[0002] CRISPR / Cas9 system, derived from the immune mechanism of bacteria to degrade invading viral DNA or other foreign DNA. Using RNA-mediated DNA-binding and endonuclease activities, this system can be regulated in the genome in a sequence-dependent manner. Cas9 protein binds and cleaves double-stranded DNA in a sequence-specific manner, and this sequence-specific binding is achieved through a guide RNA (gRNA) complementary to the target sequence and the adjacent protospacer-adjacent motif (PAM). The complex formed by Cas9 and gRNA mediates DNA double-strand breaks, and also completes targeted gene editing through two DNA repair mechanisms, NHEJ and HDR. The CRISPR / Cas9 system can also be combined with different effector molecules to expand its functions, enabling transcripti...

Claims

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