通过显微注射未受精卵母细胞对蜥蜴进行 CRISPR-Cas9 基因编辑。
CRISPR-Cas9 Gene Editing in Lizards through Microinjection of Unfertilized Oocytes.
机构信息
Department of Genetics, University of Georgia, Athens, GA 30602, USA; Department of Cellular Biology, University of Georgia, Athens, GA 30602, USA.
Department of Genetics, University of Georgia, Athens, GA 30602, USA.
出版信息
Cell Rep. 2019 Aug 27;28(9):2288-2292.e3. doi: 10.1016/j.celrep.2019.07.089.
CRISPR-Cas9-mediated gene editing has enabled the direct manipulation of gene function in many species. However, the reproductive biology of reptiles presents unique barriers for the use of this technology, and there are no reptiles with effective methods for targeted mutagenesis. Here, we demonstrate that the microinjection of immature oocytes within the ovaries of Anolis sagrei females enables the production of CRISPR-Cas9-induced mutations. This method is capable of producing F0 embryos and hatchlings with monoallelic or biallelic mutations. We demonstrate that these mutations can be transmitted through the germline to establish genetically modified strains of lizards. Direct tests of gene function can now be performed in Anolis lizards, an important model for studies of reptile evolution and development.
CRISPR-Cas9 介导的基因编辑技术使许多物种的基因功能直接操纵成为可能。然而，爬行动物的生殖生物学为这项技术的应用带来了独特的障碍，目前还没有具有有效靶向诱变方法的爬行动物。在这里，我们证明了将不成熟的卵母细胞微注射到雌性 Anolis sagrei 的卵巢内，可产生 CRISPR-Cas9 诱导的突变。该方法能够产生具有单等位基因突变或双等位基因突变的 F0 胚胎和幼体。我们证明这些突变可以通过生殖系传递，从而建立遗传修饰的蜥蜴品系。现在可以在 Anolis 蜥蜴中直接进行基因功能测试，这是研究爬行动物进化和发育的重要模型。
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