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CRISPR-Cas9-Based Genome Editing in the Silverleaf Whitefly (Bemisia tabaci)

机译:银叶粉虱(Bemisia tabaci)中基于CRISPR-Cas9的基因组编辑

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摘要

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.
机译:中东亚小I(MEAM1)属隐性物种是一种严重的农业多食性害虫,是多种植物病毒的媒介,在世界范围内造成重大经济损失。由于缺乏健壮的基因编辑工具,控制受到限制。由于小胚胎在技术上具有挑战性,并且注射后死亡率高,因此难以进行基因编辑。我们开发了一种基于注射卵黄成年雌性而不是胚胎的CRISPR-Cas9基因编辑方案(“ ReMOT Control”)。我们鉴定了卵巢靶向肽配体(“ BtKV”),当与Cas9融合并注入成年雌性后,将核糖核蛋白复合物转导至种系,从而有效,遗传地编辑了后代基因组。与胚胎注射相反,成人注射很容易,不需要专门的设备。开发易于使用的基因编辑协议将使研究人员能够将逆向遗传方法应用于该物种,并为这种毁灭性害虫带来新的防治方法。

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