Construction and expression verification of mouse spermatogenic cell-specific expression vector AAV-Dazl-RFP-Flag

Abstract

Abstract: Objective To study the effect of spermatogenic cell-specific promoter Dazl in Spermatogenic cells. Constructing the eukaryotic reporter gene expression vector AAV-Dazl-RFP-Flag containing spermatogenic cell-specific promoter Dazl. Methods The Adeno-associated virus vector AAV-CMV-RFP-Flag was modified by homologous recombination to construct mouse permatogenic cell-specific expression vector AAV-Dazl-RFP-Flag. In vitro, GC1-spg wild-type cells and HEK-293T cells were transfected, and the expression specificity of AAV-Dazl-RFP-Flag in vitro was verified by immunofluorescence, q-PCR, and Western blot. In vivo, AAV-Dazl-RFP-Flag was packaged with adeno-associated virus (AAV), and AAV was injected into the testes of mouse through microinjection technology, then the expression specificity of AAV-Dazl-RFP-Flag in vivo was detected by q-PCR. Results Mouse Sper- matogenic cell-specific expression vector AAV-Dazl-RFP-Flag is specifically expressed in spermatogenic, cells thus provid providing a more efficient tool for gene therapy of male reproductive infertility.

Key words: Dazl, promoter, gene therapy, spermatogenesis

CLC Number:

R334

LI Ya-hui, LI Kai, LUO Yan-yun, LI Meng-zhen, MANG Xin-yu, SONG Wei, YANG Tao. Construction and expression verification of mouse spermatogenic cell-specific expression vector AAV-Dazl-RFP-Flag[J]. Basic & Clinical Medicine, 2020, 40(6): 753-758.

References

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