Department of Entomology, The Pennsylvania State University, University Park, Pennsylvania, USA.
Department of Entomology, Cornell University, Ithaca, New York.
CRISPR J. 2020 Apr;3(2):89-96. doi: 10.1089/crispr.2019.0067.
cryptic species Middle East-Asia Minor I (MEAM1) is a serious agricultural polyphagous insect pest and vector of numerous plant viruses, causing major worldwide economic losses. control is limited by lack of robust gene editing tools. Gene editing is difficult in due to small embryos that are technically challenging to inject and which have high mortality post injection. We developed a CRISPR-Cas9 gene editing protocol based on injection of vitellogenic adult females rather than embryos ("ReMOT Control"). We identified an ovary-targeting peptide ligand ("BtKV") that, when fused to Cas9 and injected into adult females, transduced the ribonucleoprotein complex to the germline, resulting in efficient, heritable editing of the offspring genome. In contrast to embryo injection, adult injection is easy and does not require specialized equipment. Development of easy-to-use gene editing protocols for will allow researchers to apply the power of reverse genetic approaches to this species and will lead to novel control methods for this devastating pest insect.
中东-亚洲小种 1 号（MEAM1）是一种严重的农业多食性害虫和许多植物病毒的载体，给全世界造成重大经济损失。由于缺乏强大的基因编辑工具，因此对其进行控制受到限制。由于胚胎小，技术上难以注射，且注射后死亡率高，因此在 中进行基因编辑很困难。我们开发了一种基于注射卵黄生成期成年雌性而不是胚胎的 CRISPR-Cas9 基因编辑方案（“ReMOT Control”）。我们鉴定了一种卵巢靶向肽配体（“BtKV”），当它与 Cas9 融合并注射到成年雌性体内时，会将核糖核蛋白复合物转导到生殖系，从而使后代基因组得到高效、可遗传的编辑。与胚胎注射相比，成年注射简单，不需要专门的设备。为 开发易于使用的基因编辑方案将使研究人员能够将反向遗传方法的力量应用于该物种，并为这种破坏性害虫提供新的控制方法。
