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sgRNA targeting sequence specifically targeting mouse Krt14 gene and application thereof

A species-specific, mouse-based technology, applied in the fields of medical genetics and molecular biology, can solve problems such as the lack of sgRNA guide sequences, and achieve the effects of efficient screening, high concentration, and increased quantity

Active Publication Date: 2020-02-11
人科(北京)生物技术有限公司
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no sgRNA guide sequence for the mouse Krt14 gene and no method to edit the gene

Method used

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  • sgRNA targeting sequence specifically targeting mouse Krt14 gene and application thereof
  • sgRNA targeting sequence specifically targeting mouse Krt14 gene and application thereof
  • sgRNA targeting sequence specifically targeting mouse Krt14 gene and application thereof

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Embodiment Construction

[0083] The principles and features of the present invention will be described below in conjunction with specific drawings, and the examples given are only used to explain the present invention, not to limit the scope of the present invention.

[0084] 1. The human KRT14 gene pathogenic mutation site was screened, and the human KRT14 p.Arg125 site was identified as the pseudo-mutation site.

[0085] Epidermolysis bullosa simplex is caused by mutations in the KRT14 gene. First, the pathogenic mutations in the human KRT14 gene were screened using the OMIM database, and a total of 19 mutation sites were screened (as shown in Table 1); Extensive screening of the pathogenic mutation of KRT14, the present invention also utilizes the ExAC online database to retrieve 9 kinds of KRT14 function inactivation mutations (such as figure 1 and shown in Table 2); then, the present invention utilizes the ClinVar database to retrieve the KRT14 pathogenic mutation situation again, retrieves 36 ki...

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Abstract

The invention discloses a sgRNA targeting sequence specifically targeting a mouse Krt14 gene and application thereof, and belongs to the technical fields of medical genetics and molecular biology. A nucleotide sequence corresponding to sgRNA is any one of four sequences of SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4. The invention further discloses a method for editing the mouse Krt14 gene by using the sgRNA targeting sequencing specifically targeting the mouse Krt14 gene. The sgRNA targeting sequence can mediate the efficient cleavage of target DNA by Cas9 protein, and then be usedfor editing the mouse Krt14 gene to affect the function of protein encoded by the mouse Krt14 gene. The sgRNA targeting sequence can achieve efficient targeting by a CRISPR / Cas9 system, and the efficiency is 70%, 90%, 65% and 100% separately.

Description

technical field [0001] The invention relates to a sgRNA guide sequence specifically targeting mouse Krt14 gene and application thereof, belonging to the technical fields of medical genetics and molecular biology. Background technique [0002] The CRISPR / Cas system was first discovered in bacteria, and it performs acquired immune functions in eubacteria and archaea, which can resist the invasion of foreign viruses and plasmids. People have modified the CRISPR / Cas system of the microorganism itself by means of genetic engineering, thereby creating a targeting system that can be widely used in gene editing of higher organisms, the CRISPR / Cas9 system. Since the system came out in 2012, it has injected strong impetus into life science and biomedical research, and has become a research hotspot in recent years. Since 2013, researchers have successfully edited the genome in mammalian cells by using the DNA binding activity and endonuclease activity of CRISPR / Cas9. In the CRISPR / Ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/85C12N15/90C12N5/10
CPCC07K14/47C12N15/113C12N15/907C12N2310/20
Inventor 岳鹏鹏于鸿浩付灿李勇马海林
Owner 人科(北京)生物技术有限公司
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