The Application Of CRISPR/Cas9 In Mouse In Vivo Electroporation

Nervous system is very important for higher organisms.However,there are a lot of mysteries about the genes that related to neurogenesis and development to be discovered.Here,we discribed a method to study genes' function in neurogenesis and development.By applying CRISPR/Cas9 in mouse in vivo electroporation,we found CRISPR/Cas9 has high efficiency in knocking down and overexpressing genes in the nervous system.We first validated the function of DCX.DCX is very important for neuron migration,and its knockdown leads to neuron migration defects.This proved the correctness of our conjecture and then we used this method to study the function of constant exons of Pcdh? cluster.We found that after deleting constant exons of Pcdh? cluster,neurons' migration are apparently defective,suggesting that the constant exons of Pcdh? cluster are very important for neuron migration.Besides knocking down genes,we used CRISPR/Cas9 system to overexpress Pyk2,to explore whether this way would work to overexpress genes in mouse in vivo electroporation.The results turned out to be positive.Finally,the study of Pcdh? cluster regulation region HS5-1 proved that this method can be used to research the function of gene regulation region.To further know the mechanism of Cas9 in nervous cells,we designed experiments to identify the genotype of target genes.The results showed that applying CRISPR/Cas9 in mouse in vivo electroporation will lead to efficient deletion,inversion and duplication of target genes.These results are both verified in gene and gene regulation regions.This study provides a method to research the genes functioning in neurogenesis and development,and at the same time proves that in mouse in vivo electroporation will lead to efficient deletion,inversion and duplication of target genes.