SRY is the Y-chromosomal gene that determines male sex development in humans and most other mammals. After three decades of study, we still lack a detailed understanding of which domains of the SRY protein are required to engage pathway of gene activity leading to testis development. Some insight has been gained from the study of genetic variations underlying differences/disorders of sex determination (DSD), but the lack of a system of experimentally generating SRY mutations and studying their consequences in vivo has limited progress in the field. To address this issue, we generated a mouse model carrying a human SRY transgene able to drive male sex determination in XX mice. Using CRISPR-Cas9 gene editing, we generated novel genetic modifications in each of SRYs three domains (N-terminal, HMG box, and C-terminal) and performed detailed analysis of their molecular and cellular effects on embryonic testis development. Our results provide new functional insights unique to human SRY and the causes of DSD, and present a versatile and powerful system in which to demonstrate causality of SRY variations in DSD, to functionally study the SRY variation database, and to characterize new pathogenic SRY variations found in DSD.

中文翻译：

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人SRY转基因小鼠模型的产生和突变分析

SRY是决定人类和大多数其他哺乳动物中男性性别发育的Y染色体基因。经过三十年的研究，我们仍然缺乏对SRY蛋白的哪些结构域参与导致睾丸发育的基因活性途径的详细了解。从性别差异/性别决定（DSD）的潜在遗传变异研究中获得了一些见识，但是缺乏通过实验产生SRY突变并在体内研究其后果的系统，在该领域的进展有限。为了解决这个问题，我们生成了一个带有人类SRY转基因的小鼠模型，该模型能够驱动XX小鼠中的男性性别决定。使用CRISPR-Cas9基因编辑，我们在SRY的三个域（N端，HMG框，和C末端），并对其分子和细胞对胚胎睾丸发育的影响进行了详细分析。我们的结果为人类SRY和DSD的成因提供了独特的新功能见解，并提供了一种通用而强大的系统，可在其中证明DSD中SRY变异的因果关系，在功能上研究SRY变异数据库并表征发现的新病原性SRY变异在DSD中。