Method for knocking out two mir-505 alleles

A biallelic, gene technology, applied in DNA/RNA fragments, introduction of foreign genetic material using vectors, recombinant DNA technology, etc., can solve problems such as unclear specific methods
CN104651401AInactive Publication Date: 2015-05-27DONGHUA UNIV
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Patent Information

Authority / Receiving Office
CN ยท China
Patent Type
Applications(China)
Current Assignee / Owner
DONGHUA UNIV
Publication Date
2015-05-27
Estimated Expiration
Not applicable ยท inactive patent

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Abstract

The invention relates to a method for knocking out two mir-505 alleles. The method comprises the following steps: constructing two targeting vectors for mir-505 alleles, replacing and knocking one mir-505 allele out of a cell of a mammal by mediating one targeting vector by using a CRISPR / Cas system, obtaining the cell with one mir-505 allele being knocked out by adopting a G418 resistance screening method, knocking another mir-505 allele out of the cell of the mammal by mediating another green fluorescent targeting vector by using the CRISPR / Cas system based on the cells, and obtaining the cell with two mir-505 alleles being knocked out by adopting a fluorescence screening method. The method is helpful to constructing a gene knocked-out cell model and has good application prospect.
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Description

technical field

[0001] The invention belongs to the field of gene knockout, in particular to a method for knocking out mir-505 biallelic genes. Background technique

[0002] In 1987, the Japanese research group discovered a tandem spaced repeat sequence near the alkaline phosphatase gene of E.coli K12 [Ishino Y, Shinagawa H, Makino K, et al. Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene produce.J Bacteriol,1987,169(12):5429-5433], in subsequent studies, it was found that such spacer repeat sequences widely exist in the genomes of bacteria and archaea, in 2002 In 2010, it was officially named by scientists as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)[Jansen R, van Embden J D, Gaastra W, et al. Identification of a novel family of sequence repeats among prokaryotes.Omics: a journal of Intearative Biology, 2002, 6(1): 23-33], [Mojica F J, Ferrer C, Juez G,...

Claims

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