Fusion protein in Cpf1 and p300 core structural domain, corresponding DNA target activation system and application

Abstract

The invention discloses a fusion protein in a Cpf1 and p300 core structural domain, a corresponding DNA target activation system and application. CRISPR / Cpf1 with enzyme cutting activity is mutated to be free of enzyme cutting activity and of target gene identification characteristic, is fused with p300 protein transcriptional activation, reaches the purpose of target gene activation inside mammalian cells, and has the characteristics of being simple, efficient and high in specificity. The fusion protein has the advantages of low off-target rate, cost saving and the like. The system can be mutually complementary with an existing target gene activation method based on CRISPR / Cas9, the gene editing range of the CRISPR / Cas9 system is expanded, and the system has a good application prospect.

Description

technical field [0001] The invention relates to the fields of genetic engineering and biotechnology, in particular to the targeted gene activation of mammalian cells by using CRISPR / Cpf1 and its variants and p300 fusion protein. Background technique [0002] CRISPR / Cas (Clustered Regularly Interspaced Short Palindromic Repeats / CRISPR-Associated systems), the full name is the constant palindromic repeat sequence cluster / constant palindromic repeat sequence cluster-associated protein system, which is an acquisition of bacteria and archaea immune system. In 2010, Garneau et al. found that Cas9 is the only Cas nuclease that can mediate DNA cleavage in the CRISPR type II system of Streptococcus thermophilus. In 2013, Zhang Feng and other scientists first reported the application of CRISPR-Cas9 system in mammalian genome editing, which opened up the powerful gene editing function of CRISPR-Cas9 and became a new star in the field of life sciences, which has been widely studied and...

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to mutate the CRISPR / Cpf1 with enzymatic cleavage activity into a targeted gene with no enzymatic cleavage activity in view of the problems of low targeting, low activation efficiency, and complex action systems involving the existing gene activation methods. Identify the characteristics and integrate the transcriptional activation of p300 protein to provide a fusion protein, expression vector and corresponding DNA-targeted activation system that can target gene activation in mammalian cells

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