Method for increasing non-homologous end joining efficiency of CRIPSR/Cas9 target knock-out genes

A gene and efficiency technology, applied in the field of improving the efficiency of non-homologous end joining of CRIPSR/Cas9 targeted knockout genes, can solve the problems of reducing the fidelity of the repair process and gene breakage
CN106906242AInactive Publication Date: 2017-06-30重庆高圣生物医药有限责任公司
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
重庆高圣生物医药有限责任公司
Publication Date
2017-06-30
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention belongs to the technical field of molecular biology and biomedicine, and relates to a method for increasing the efficiency of CRIPSR / Cas9 target knock-out genes NHEJ. According to the method for increasing the efficiency of CRIPSR / Cas9 target knock-out gene NHEJ, small molecule interference RNA that efficiently interferes with expression of PTEN genes is screened out; by using CRISPR / Cas9 system to target knockout genes, the small molecule RNA is cotransfected into the target gene, thus the NHEJ efficiency of target genes is effectively improved and the NHEJ of target gene in cell multiple targets is significantly improved. The method has the advantages of low cost, simple operation and high efficiency, and has a good promoting effect on the efficiency and application of CRISPR / Cas9 technology.
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Description

technical field

[0001] The invention belongs to the technical fields of molecular biology and biomedicine, and in particular relates to a method for improving the efficiency of non-homologous end joining (NHEJ) of CRIPSR / Cas9 targeted knockout genes. Background technique

[0002] The CRISPR / Cas system is developed from the adaptive immune system of bacteria and archaea against foreign viruses or plasmids, including three different types, of which the DNA endonuclease Cas9 of the Type Ⅱ CRISPR / Cas system has only one subclass. The base has the simplest structure, so it is the most widely used. In addition to the Cas9 protein, the system also includes two short CRISPR RNAs (crRNAs) and trans-activating crRNAs (tracrRNA). The mature crRNA-tracrRNA complex can guide the Cas9 protein to the target sequence through complementary base pairing, and specifically cut the DNA double strand near the PAM (protospace radjacent motif) to form a DSB (double strand break). DSB can be repai...

Claims

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