Advanced Science. 2023 Sep 11
Kai Xia, Fulin Wang, Zhipeng Tan, Suyuan Zhang, Xingqiang Lai, Wangsheng Ou, Cuifeng Yang, Hong Chen, Hao Peng, Peng Luo, Anqi Hu, Xiang’an Tu, Tao Wang, Qiong Ke, Chunhua Deng, Andy Peng Xiang
Products used in the paper Details Operation
Lentivirus packaging WT-SLCs at passage 1 were transduced with mCherry lentivirus (PackGene Biotech, Guangzhou, China) at 30% confluence in 6-well plates.  Request Quote

Research Field: testis

Routes of Administration: into testis or oocytes

Targeted organ: testis

Animal or cell line strain: Mouse

Abstract

Hereditary primary hypogonadism (HPH), caused by gene mutation related to testosterone synthesis in Leydig cells, usually impairs male sexual development and spermatogenesis. Genetically corrected stem Leydig cells (SLCs) transplantation may provide a new approach for treating HPH. Here, a novel nonsense-point-mutation mouse model (LhcgrW495X) is first generated based on a gene mutation relative to HPH patients. To verify the efficacy and feasibility of SLCs transplantation in treating HPH, wild-type SLCs are transplanted into LhcgrW495X mice, in which SLCs obviously rescue HPH phenotypes. Through comparing several editing strategies, optimized PE2 protein (PEmax) system is identified as an efficient and precise approach to correct the pathogenic point mutation in Lhcgr. Furthermore, delivering intein-split PEmax system via lentivirus successfully corrects the mutation in SLCs from LhcgrW495X mice ex vivo. Gene-corrected SLCs from LhcgrW495X mice exert ability to differentiate into functional Leydig cells in vitro. Notably, the transplantation of gene-corrected SLCs effectively regenerates Leydig cells, recovers testosterone production, restarts sexual development, rescues spermatogenesis, and produces fertile offspring in LhcgrW495X mice. Altogether, these results suggest that PE-based gene editing in SLCs ex vivo is a promising strategy for HPH therapy and is potentially leveraged to address more hereditary diseases in reproductive system.

Popular Services

AAV Packaging Service

AAV Analytical Service

Vector Design