CRISPR/Cas9 mediated knockout of the abdominal-A homeotic gene in the global pest, diamondback moth (Plutella xylostella)
CRISPR/Cas9 mediated knockout of the abdominal-A homeotic gene in the global pest, diamondback moth (Plutella xylostella)
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CRISPR/Cas9 介导敲除全球害虫小菜蛾 (Plutella xylostella) 中的腹部 A 同源基因
DOI：
10.1016/j.ibmb.2016.06.004
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发表时间：
2016-08-01
影响因子：
3.8
通讯作者：
You, Minsheng
中科院分区：
文献类型：
--
作者：
Huang, Yuping;Chen, Yazhou;You, Minsheng
小菜蛾 (Plutella xylostella (L.)) 是一种世界性农业害虫，已对多种杀虫剂产生抗药性。基于遗传学的方法显示出作为替代害虫管理方法的前景，但需要功能研究来确定合适的基因目标。在这里，我们使用 CRISPR/Cas9 系统来靶向腹部-A 基因，该基因在确定腹部节段的身份和功能方面发挥着重要作用。我们报道，小菜蛾腹部-A (Pxabd-A) 具有两种结构相似的剪接异构体（A 和 B），仅外显子 II 的长度不同，异构体 A 中多了 15 个核苷酸。在所有发育阶段都检测到了 Pxabd-A 转录本，特别是在蛹和成虫中。注射 448 个卵子后，基于 CRISPR/Cas9 的 Pxabd-A 外显子 I 诱变产生了 91% 的嵌合突变体。在孵化的幼虫和未孵化的胚胎中可以看到具有异常前肢和畸形节段的表型，并且各种缺陷被遗传给下一代（G（1））。突变体的基因分型表明 Pxabd-A 基因组位点存在多个突变。结果表明，不仅在Go幸存者中，而且在G1个体中，Pxabd-A位点诱导了一系列插入和缺失，这为基因组编辑提供了基础。我们的研究证明了 CRISPR/Cas9 系统在农业害虫中靶向基因的实用性，因此为开发新型害虫管理工具奠定了基础。 (C) 2016 Elsevier Ltd. 保留所有权利。
The diamondback moth, Plutella xylostella (L.), is a worldwide agricultural pest that has developed resistance to multiple classes of insecticides. Genetics-based approaches show promise as alternative pest management approaches but require functional studies to identify suitable gene targets. Here we use the CRISPR/Cas9 system to target a gene, abdominal-A, which has an important role in determining the identity and functionality of abdominal segments. We report that P. xylostella abdominal-A (Pxabd-A) has two structurally-similar splice isoforms (A and B) that differ only in the length of exon II, with 15 additional nucleotides in isoform A. Pxabd-A transcripts were detected in all developmental stages, and particularly in pupae and adults. CRISPR/Cas9-based mutagenesis of Pxabd-A exon I produced 91% chimeric mutants following injection of 448 eggs. Phenotypes with abnormal prolegs and malformed segments were visible in hatched larvae and unhatched embryos, and various defects were inherited by the next generation (G(1)). Genotyping of mutants demonstrated several mutations at the Pxabd-A genomic locus. The results indicate that a series of insertions and deletions were induced in the Pxabd-A locus, not only in Go survivors but also in G1 individuals, and this provides a foundation for genome editing. Our study demonstrates the utility of the CRISPR/Cas9 system for targeting genes in an agricultural pest and therefore provides a foundation the development of novel pest management tools. (C) 2016 Elsevier Ltd. All rights reserved.