Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, 130062, China.
CAS Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530, China.
Sci Rep. 2017 May 8;7(1):1569. doi: 10.1038/s41598-017-01727-y.
The 3' untranslated regions (UTRs), located at the end of mRNA molecules, are believed to play a role in RNA replication and/or protein translation. Mutations in the tyrosinase (Tyr) gene are known to cause recessive albinism in humans and other species. In this study, to test whether the CRISPR/Cas9 system works on the mutation of the UTRs regulatory region in rabbit, the 3' UTR of the rabbit Tyr gene was deleted by a dual sgRNA directed CRISPR/Cas9 system. As expected, gray coat color and reduced melanin in hair follicles and irises was found in the mutated rabbit, thus increasing confidence in the association of the mutation of the Tyr 3' UTR with graying in rabbit. The graying phenotype was also found in the F1 generation, suggesting that the mutated allele can be stably inherited by the offspring. Thus, we provide the first evidence that reduced melanin and graying can be caused by deletion of the Tyr 3' UTR in rabbits. Additionally, CRISPR/Cas9-mediated large fragment deletions can facilitate genotype to phenotype studies of UTRs or non-coding RNAs in future.
3'非翻译区（UTR）位于 mRNA 分子的末端，被认为在 RNA 复制和/或蛋白质翻译中发挥作用。已知酪氨酸酶（Tyr）基因的突变会导致人类和其他物种的隐性白化病。在这项研究中，为了测试 CRISPR/Cas9 系统是否适用于兔UTR 调节区的突变，通过双 sgRNA 指导的 CRISPR/Cas9 系统删除了兔 Tyr 基因的 3'UTR。正如预期的那样，突变兔的毛色呈灰色，毛囊和虹膜中的黑色素减少，增加了 Tyr 3'UTR 突变与兔毛变白之间关联的可信度。这种灰色表型也出现在 F1 代中，表明突变等位基因可以稳定遗传给后代。因此，我们首次提供了证据，表明黑色素减少和白化可以通过删除兔 Tyr 3'UTR 引起。此外，CRISPR/Cas9 介导的大片段缺失可以促进未来UTR 或非编码 RNA 的基因型到表型研究。
