Fusion protein based on CjCas9 and VPR core structure domains, corresponding DNA targeting activation system and application of system

A DNA targeting and fusion protein technology, applied in recombinant DNA technology, DNA/RNA fragments, stably introducing foreign DNA into chromosomes, etc., can solve the problems of low targeting, too large system size, and low activation efficiency
CN110358753AActive Publication Date: 2019-10-22SOUTHERN MEDICAL UNIVERSITY
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Patent Information

Authority / Receiving Office
CN ยท China
Current Assignee / Owner
SOUTHERN MEDICAL UNIVERSITY
Publication Date
2019-10-22

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Abstract

The invention discloses a fusion protein based on CjCas9 and VPR core structure domains, a corresponding DNA targeting activation system and application of the system. The fusion protein comprises twoheterologous polypeptide structure domains, and one polypeptide structure domain comprises VPR protein with the transcription activation activity; the other polypeptide structure domain comprises theCjCas9 protein, the CjCas9 protein is in a dCjCas9 subtype or a mini-dCjCas9 subtype or a CjCas9 wild type, and the dCjCas9 subtype contains D8A and H559A single-locus amino acid mutation; the mini-dCjCas9 subtype contains D8A single-locus amino acid mutation and deficiency of most HNH structure domains. The DNA targeting activation system comprises the fusion protein and one or more kinds of guide RNA, and the guide RNA comprises a sequence with the length of 14-22 bp and a framework sequence with the length of 80 bp, wherein the sequences are designed for a target gene promoter area. The DNA targeting activation system is applied to targeting activation of a gene. The DNA targeting activation system has the advantages of being small in size, capable of achieving high specificity, a highactivation effect and easy synthesis, low in cost and the like.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, and relates to a fusion protein based on CjCas9 and its mutants and a VPR core domain, a corresponding DNA targeting activation system and applications thereof. Background technique

[0002] The CRISPR / Cas9 system is currently a widely used gene editing system, which is transformed from the adaptive immune mechanism of bacteria and archaea to degrade foreign DNA. The CRISPR / Cas9 system mainly consists of two components: Cas9 protein and guide RNA. After the combination of Cas9 protein and guide RNA, it will cut the double strand of DNA at a specific site under the guidance of guide RNA, resulting in a break, which is much simpler than the previous gene editing method. In recent years, the CRISPR / Cas9 system has also been gradually applied in the field of gene regulation. Using the CRISPR / Cas9 system that has lost its cleavage activity and combined with transcriptional activators or repressors can a...

Claims

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