A fly model establishes distinct mechanisms for synthetic CRISPR/Cas9 sex distorters
A fly model establishes distinct mechanisms for synthetic CRISPR/Cas9 sex distorters
复制标题
果蝇模型建立了合成 CRISPR/Cas9 性别扭曲剂的独特机制
DOI：
10.1101/834630
复制
发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Fasulo B
中科院分区：
文献类型：
--
作者：
Fasulo B
合成性别扭曲最近已在疟疾蚊子中开发出来，依赖于在精子发生期间靶向X染色体的核酸内切酶。虽然受到自然发生的特征的启发，但仍然不清楚它们是如何发挥作用的，并且考虑到它们的遗传控制潜力，这种策略在物种之间的可移植性如何。我们为CRISPR/Cas9性别比例扭曲的两种不同机制建立了果蝇模型-“X-粉碎”和“X-中毒”-并剖析了它们的靶位点要求和修复动力学。当Cas9核酸内切酶活性发生在精子发生的减数分裂阶段时，X-切割导致性别畸变，但当Cas9从干细胞阶段开始表达时则不会。我们的研究结果表明，X-粉碎被NHEJ DNA修复途径抵消，可以在非必需序列的单个重复簇上操作，尽管靶向一些这样的重复对性别比没有影响。相比之下，X中毒，即靶向X染色体上推定的单倍致死基因，当我们将Cas9切割引导至X连锁的核糖体靶基因RpS 6时，诱导了对雄性的高偏倚（>92%）。在X中毒的情况下，性别扭曲与生殖输出的损失相关联，尽管显性-负效应似乎驱动了雌性致死的机制。这些模型系统将指导研究和应用性别扭曲的医学或农业上重要的昆虫目标物种。
Synthetic sex distorters have recently been developed in the malaria mosquito, relying on endonucleases that target the X-chromosome during spermatogenesis. Although inspired by naturally-occurring traits, it has remained unclear how they function and, given their potential for genetic control, how portable this strategy is across species. We establishedDrosophilamodels for two distinct mechanisms for CRISPR/Cas9 sex-ratio distortion—“X-shredding” and “X-poisoning”—and dissected their target-site requirements and repair dynamics. X-shredding resulted in sex distortion when Cas9 endonuclease activity occurred during the meiotic stages of spermatogenesis but not when Cas9 was expressed from the stem cell stages onwards. Our results suggest that X-shredding is counteracted by the NHEJ DNA repair pathway and can operate on a single repeat cluster of non-essential sequences, although the targeting of a number of such repeats had no effect on the sex ratio. X-poisoning by contrast, i.e. targeting putative haplolethal genes on the X chromosome, induced a high bias towards males (>92%) when we directed Cas9 cleavage to the X-linked ribosomal target geneRpS6. In the case of X-poisoning sex distortion was coupled to a loss in reproductive output, although a dominant-negative effect appeared to drive the mechanism of female lethality. These model systems will guide the study and the application of sex distorters to medically or agriculturally important insect target species.
登录
查看更多内容
影响因子：
64.5
作者：
Zetsche B;Gootenberg JS;Abudayyeh OO;Slaymaker IM;Makarova KS;Essletzbichler P;Volz SE;Joung J;van der Oost J;Regev A;Koonin EV;Zhang F
通讯作者：
Zhang F
埃及伊蚊减数分裂驱动的细胞遗传学分析
影响因子：
1.5
作者：
M. Newton;R. Wood;D. I. Southern
通讯作者：
D. I. Southern
影响因子：
4.5
作者：
Windbichler N;Papathanos PA;Crisanti A
通讯作者：
Crisanti A
影响因子：
20.3
作者：
Devlin, Emily E.;DaCosta, Lydie;Bodine, David M.
通讯作者：
Bodine, David M.
影响因子：
5.4
作者：
Mikhaylova LM;Nurminsky DI
通讯作者：
Nurminsky DI