关键词: TALEN, 显微注射, WFDC2, HE4, 基因敲除

Abstract: Ob jective To construct HE4 （WFDC2） knockoutmice using microinjection of transcription activatorlike effector nuclease（TALEN） mRNAs.Method TALENs-mediated precise knockout site specifically targeting mouse WFDC2 gene was designed，and the correspondent vectorswere constructed.TALENs cDNA was transcribed in vitro initially，then the F0-generation m ice were obtained from the microinjection of TALENs mRNA into the C57BL/6 Jmouse zygotes.F0 mice were characterized at genomic DNA level by using genotyping，and WFDC2 knockoutmice were confirmed.Result We designed and constructed TALENs which recognized and cut the first exon of Wfdc2， and got 24 F0 m ice， among which one had frameshift mutation， demonstrating that the WFDC2-knockoutm ice were generated by TALEN technique，and we also find that the homozygous null m ice died after birth.Conclusion WFDC2 gene knockout m ice model were successfully developed by using the combination of m icroinjection and TALENs-mediated deletion of mouse WFDC2 gene.This will be a useful model to study the WFDC2's physiological function in mouse，and could also provide the insight on the involvement of WFDC2 in certain diseases such as ovarian and endometrial cancer progression

Key words: TALEN, microinjection, WFDC2, HE4, gene knockout