Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod ovaries for heritable germline gene editing.
Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod ovaries for heritable germline gene editing.
复制标题
将CRISPR-CAS9核糖核蛋白的靶向递送到节肢动物卵巢中，以进行可遗传的种系基因编辑。
DOI：
10.1038/s41467-018-05425-9
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发表时间：
2018-08-01
影响因子：
16.6
通讯作者：
Rasgon JL
中科院分区：
文献类型：
--
作者：
Chaverra-Rodriguez D;Macias VM;Hughes GL;Pujhari S;Suzuki Y;Peterson DR;Kim D;McKeand S;Rasgon JL
Cas9介导的基因编辑是解决节肢动物研究问题的强大工具。目前的方法依赖于胚胎显微注射传递Cas9核糖核蛋白(RNP)复合体，这具有挑战性，仅限于少数物种，即使在优化的类群中也效率低下。在这里，我们开发了一种名为受体介导的卵巢转导货物(远程控制)的技术，通过注射到成年雌性蚊子中将Cas9 RNP传递到节肢动物生殖系。我们鉴定了一种多肽(P2C)，它介导Cas9 RNP从雌性血淋巴转导到发育中的蚊子卵母细胞，导致后代的基因编辑可遗传，每注射一只蚊子的效率高达0.3个突变体。我们证明了P2C在六种蚊子中的功能。识别特定于分类群的卵巢特异性配体-受体对可能进一步扩大远程控制在新物种中进行基因编辑的使用。Cas9核糖核蛋白复合体的输送可以通过向卵子中微量注射来完成，尽管这在技术上是一个具有挑战性的过程。在这里，作者展示了远程控制，通过标记来自卵黄蛋白1的多肽将RNPs传递到蚊子生殖系。
Cas9-mediated gene editing is a powerful tool for addressing research questions in arthropods. Current approaches rely upon delivering Cas9 ribonucleoprotein (RNP) complex by embryonic microinjection, which is challenging, is limited to a small number of species, and is inefficient even in optimized taxa. Here we develop a technology termed Receptor-Mediated Ovary Transduction of Cargo (ReMOT Control) to deliver Cas9 RNP to the arthropod germline by injection into adult female mosquitoes. We identify a peptide (P2C) that mediates transduction of Cas9 RNP from the female hemolymph to the developing mosquito oocytes, resulting in heritable gene editing of the offspring with efficiency as high as 0.3 mutants per injected mosquito. We demonstrate that P2C functions in six mosquito species. Identification of taxa-specific ovary-specific ligand–receptor pairs may further extend the use of ReMOT Control for gene editing in novel species. Delivery of Cas9 ribonucleoprotein complexes can be done via microinjection into eggs, though this is a technically challenging procedure. Here the authors demonstrate ReMOT Control, delivery of RNPs to the mosquito germline by tagging with peptide derived from yolk protein 1.
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影响因子：
4.2
作者：
Gilabert-Oriol, Roger;Thakur, Mayank;Weng, Alexander
通讯作者：
Weng, Alexander
DOI：
10.1073/pnas.1711538114
发表时间：
2017-12-05
影响因子：
11.1
作者：
Li M;Bui M;Yang T;Bowman CS;White BJ;Akbari OS
通讯作者：
Akbari OS
影响因子：
64.8
作者：
Catteruccia, F;Nolan, T;Crisanti, A
通讯作者：
Crisanti, A
影响因子：
3.8
作者：
Cheon, HM;Seo, SJ;Raikhel, AS
通讯作者：
Raikhel, AS
影响因子：
2.4
作者：
Kiesgen, Stefan;Liebers, Nora;Krauss, Juergen
通讯作者：
Krauss, Juergen