Method of introducing random insertion mutation to genome of in-vivo spermatogonial stem cells

Abstract

The invention belongs to the technical field of biology and relates to a method of introducing random insertion mutation to genome of in-vivo spermatogonial stem cells. In the introduction of insertion mutation, provided is a non-integrated lentivirus composite carrier carrying a piggyBac transposon system and its preparation method. The carrier and the method are applicable to efficiently preparing non-human gene mutation animals. A gene mutation animal prepared by the method is applicable to animal phenotypic analysis, gene function research, Human genetic disease simulation, novel drug screening and novel drug target finding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for introducing random insertion mutations into the genome of spermatogonial stem cells in vivo. The invention can be used to efficiently prepare non-human gene mutant animals. Background technique [0002] Gene mutations can change the DNA nucleotide sequence of a specific gene, and specific types include base substitution mutations, frameshift mutations, deletion mutations, and insertion mutations. Gene mutations cause changes in the expression or function of specific gene products, resulting in changes in biological processes at the cellular level and abnormal phenotypes at the animal individual level ("Introduction to Genetic Analysis" 10th edition edited by Griffiths et al.). [0003] Experimental animal models carrying genetic mutations are very important resources in biological research and new drug development. In biological research, animals carrying random genetic ...

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Problems solved by technology

However, in terms of inducing gene mutations, since the integrated lentivirus prefers to insert into the transcriptionally active region of the genome, it cannot completely mediate random insertion covering the whole genome (Schroder et al., 2002, Cell110:521-529; Mitchell et al., 2004PLoSbiology2; e234)

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