Single cell RNA-sequencing of testes from control and Brca1 germ cell-specific knockout mice

To examine the role of BRCA1 during the formation of undifferentiated spermatogonia, we generated Brca1 germ cell-specific knockout mice, in which BRCA1 was undetectable when undifferentiated spermatogonia started to form at PD1. To analyze the defects comprehensively, we isolated testes from one pair of control and Brca1 germ cell-specific knockout mice male mice at PD7 and performed single-cell RNA sequencing (scRNA-seq) analyses using the 10 Genomics platform. In the study, we found that BRCA1 loss indeed disrupts early postnatal germ cell development. A pair of PD7 testes from different mice were isolated and single cells were generated for the downstream RNA-sequencing为探究乳腺癌易感基因1（BRCA1）在未分化精原细胞（undifferentiated spermatogonia）的形成过程中所发挥的作用，我们构建了生殖细胞特异性敲除Brca1的小鼠模型，该模型在出生后第1天（PD1）未分化精原细胞开始形成时，BRCA1即无法被检测到。为全面分析该敲除模型的表型缺陷，我们于出生后第7天（PD7）分别获取一对野生型对照与生殖细胞特异性Brca1敲除雄性小鼠的睾丸组织，并采用10 Genomics平台开展单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）分析。本研究发现，BRCA1的缺失确实会干扰出生后早期生殖细胞的发育进程。我们分离了两只不同小鼠的PD7时期睾丸组织，制备单细胞悬液用于后续RNA测序实验。