Wnt16 gene deletion type zebra fish

A gene deletion, zebrafish technology, applied in the field of gene knockout, can solve the problems of low efficiency of targeting technology and high off-target rate

Inactive Publication Date: 2016-12-07
HUNAN NORMAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

Traditional gene targeting technology is based on embryonic stem cells (ESC) and homologous recombination technology, so the efficiency of targeting technology is extremely low
At the beginning of 2013, a new artificial endonuclease clustered regularly interspacedshortpalindromic repeats (CRISPR)/CRISPR-associated (Cas) 9

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  • Wnt16 gene deletion type zebra fish
  • Wnt16 gene deletion type zebra fish
  • Wnt16 gene deletion type zebra fish

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Embodiment Construction

[0084] The present invention will be further described below in conjunction with accompanying drawing:

[0085] Include the following steps:

[0086] Step 1: CRISPR / Cas9 gene knockout target site design

[0087] Query the genomic DNA sequence and functional domain of the zebrafish wnt16 gene on the GenBankTM, Ensembl or UCSC databases, and design a pair of target sites for the wnt16 gene according to the principle of CRISPR / Cas knockout. The selection of the target sites must follow the 5'- 20bp-NGG-3' standard: the GG dinucleotide at the 5' end is part of the T7 promoter, and there is no such restriction when designing the target site, but it must be ensured that the 3' end of the target site is NGG, and the target site The selection must ensure that the insertion or deletion of bases at the target site can affect the entire domain of the wnt16 gene, thereby altering gene expression,

[0088] Two pairs of specific PCR primers are as follows:

[0089] F1 (target site a forw...

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Abstract

The invention discloses a wnt16 gene deletion type zebra fish. Compared with the prior art, specific genes in the genome of a living body can be muted more efficiently and precisely; moreover, the manufacturing is simple, the cost is lower, multiple sites on target genes can be sheared at the same time, and any number of single gene can be muted. The wnt16 gene deletion type zebra fish can be applied to research related with gene and skeleton growth and other research for finding whether deletion of wnt16 gene is related with growth of other organs such as heart or not, and thus the wnt16 gene deletion type zebra fish has a good medical research value. At the same time, the growth period of zebra fishes without wnt16 gene is obviously shortened, and the wnt16 gene deletion type zebra fish also has a good commercial value therefore.

Description

technical field [0001] The invention relates to the technical field of gene knockout, in particular to a wnt16 gene-deleted zebrafish. Background technique [0002] The Wnt gene is located at human 12q13 and encodes a transcription factor of 356 amino acids. It is generally believed that the Wnt signal transduction pathway is divided into canonical Wnt signaling pathway and non-canonical Wnt signaling pathway. The canonical Wnt signaling pathway is also known as the Wnt / p-catenin signaling pathway. The molecular mechanism of Wnt / p-catenin signal transduction is highly conserved in different species. Wnt16 is a non-canonical wnt ligand. It is generally believed that this gene mediates the β-catenin signaling pathway, can negatively regulate RANK signaling, inhibit the formation of osteoclasts, and participate in processes such as bone formation and bone resorption. Through gene differential expression profile analysis and genome association analysis, it was found that Wnt1...

Claims

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Application Information

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IPC IPC(8): C12N15/85A01K67/027
CPCC12N15/85A01K67/0276A01K2217/075A01K2227/40C12N2800/106C12N2800/80
Inventor 陈湘定刘薇薇苏幸邵梦思谭丽君邓红文
Owner HUNAN NORMAL UNIVERSITY
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