A systematic knowledge of the roles of DNA repair genes at the level of the organism has been limited due to the lack of appropriate experimental techniques. Here, we generated zebrafish loss-of-function mutants for 32 DNA repair and replication genes through multiplexed CRISPR/Cas9-mediated mutagenesis. High-throughput phenotypic characterization of our mutant collection revealed that three genes (atad5a, ddb1, pcna) are essential for proper embryonic development and hematopoiesis; seven genes (apex1, atrip, ino80, mre11a, shfm1, telo2, wrn) are required for growth and development during juvenile stage and six genes (blm, brca2, fanci , rad51, rad54l, rtel1) play critical roles in sex development. Furthermore, mutation in six genes (atad5a, brca2, polk, rad51, shfm1, xrcc1) displayed hypersensitivity to DNA damage agents. Further characterization of atad5a-/- mutants demonstrate that Atad5a is required for normal brain development and hematopoiesis. Our zebrafish mutant collection provides a unique resource for understanding of the roles of DNA repair genes at the organismal level.

中文翻译：

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斑马鱼CRISPR修复基因的CRISPR突变体的高通量产生和表型表征

由于缺乏适当的实验技术，对DNA修复基因在生物体中的作用的系统知识受到了限制。在这里，我们通过CRISPR / Cas9介导的多重诱变生成了用于32个DNA修复和复制基因的斑马鱼功能丧失突变体。我们的突变体集合的高通量表型特征表明，三个基因（atad5a，ddb1，pcna）对于适当的胚胎发育和造血至关重要。在幼年期生长发育需要七个基因（apex1，atrip，ino80，mre11a，shfm1，telo2，wrn），六个基因（blm，brca2，fanci，rad51，rad54l，rtel1）在性发育中起关键作用。此外，六个基因（atad5a，brca2，polk，rad51，shfm1，xrcc1）的突变显示出对DNA损伤剂的超敏性。atad5a-/-突变体的进一步表征表明，Atad5a是正常大脑发育和造血作用所必需的。我们的斑马鱼突变体集合为了解DNA修复基因在机体水平上的作用提供了独特的资源。