Method for increasing growth speed of zebra fish through gene knockout technology

Abstract

The invention belongs to the technical field of gene engineering, and discloses a method for increasing the growth speed of zebra fish through a gene knockout technology. The method for increasing thegrowth speed of zebra fish through the gene knockout technology comprises the following steps: searching mRNA sequences and genome sequences of actRIIB genes and alk4 genes of zebra fish through an NCBI online database; designing the knockout target sites of the actRIIB gene and the alk4 gene; carrying out in vitro transcription to obtain Cas9 mRNA; carrying out micro co-injection on the Cas9 mRNA, the actRIIB and the alk4 gene gRNA; screening F1-generation heterozygous mutation individuals and determining mutation types; obtaining F2-generation homozygous mutant individuals, and after the F1-generation heterozygous mutant individuals are mature, selfing the F1-generation individuals to obtain F2-generation individuals; and screening homozygous mutant individuals from F2 according to theMendel's law. According to the invention, co-knockout of actRIIB and alk4 of zebra fish can cause rapid growth of zebra fish.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a method for increasing the growth rate of zebrafish through gene knockout technology. Background technique [0002] Currently, the closest prior art: [0003] Improving the growth rate of fish can not only shorten the time to market, increase the breeding output per unit time, but also reduce the incidence of fish disease to a certain extent, thereby reducing the risk of breeding. The speed of fish growth is directly related to its breeding promotion and breeding benefits. Therefore, how to increase the growth speed of fish has always been the focus and hot spot of fish-related research. At present, the development technology of fast-growing transgenic fish is mostly gene overexpression technology, and the selected target genes are all growth hormone genes, such as the transgenic Yellow River carp developed by Academician Zhu Zuoyan's team and the transg...

AI Technical Summary

Problems solved by technology

[0006] To sum up, the problem of existing technology is that the research on actRIIB and alk4 in fish is still in its infancy

[0008]1 The efficiency of using CRIPR / Cas9 technology to knock out two genes at the same time is relatively low, and a large number of F1 individuals need to be screened

[0009]2 requires 3 passages to obtain homozygous individuals, and the proportion of homozygous individuals in F2 is only 6.25%, and the screening workload is relatively large

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