Sabin Christina E, Lauderdale James D, Menke Douglas B
Neuroscience Division of the Biomedical and Translational Sciences Institute, University of Georgia, Athens, Georgia 30602, USA.
Department of Cellular Biology, University of Georgia, Athens, Georgia 30602, USA.
Cold Spring Harb Protoc. 2025 Jul 31. doi: 10.1101/pdb.top108535.
lizards are an ecologically diverse group that includes more than 400 described species. These reptiles have been the subject of wide-ranging studies, from speciation and convergent evolution to climate adaptation and tail regeneration. While CRISPR-based gene editing has tremendous potential to reveal new insights into these and other aspects of biology, the reproductive biology of these reptiles has presented significant barriers to gene editing. Here, we briefly summarize gene editing approaches in vertebrates and discuss some of the major challenges associated with the performance of gene editing in anoles. We then introduce a recently established surgical procedure that enables the injection of CRISPR-Cas into the developing oocytes of female lizards. This approach circumvents the need to manipulate early-stage embryos and permits the production of gene-edited anoles. This method has recently been successfully adapted for use in other reptiles, suggesting that it may be effective in a wide range of species and will broadly enable studies of gene function in reptiles.
蜥蜴是一个生态多样的类群，包括400多种已被描述的物种。这些爬行动物一直是广泛研究的对象，从物种形成和趋同进化到气候适应和尾巴再生。虽然基于CRISPR的基因编辑在揭示生物学的这些及其他方面的新见解方面具有巨大潜力，但这些爬行动物的生殖生物学给基因编辑带来了重大障碍。在这里，我们简要总结脊椎动物中的基因编辑方法，并讨论与安乐蜥基因编辑操作相关的一些主要挑战。然后，我们介绍一种最近建立的手术程序，该程序能够将CRISPR-Cas注入雌性蜥蜴发育中的卵母细胞。这种方法避免了操纵早期胚胎的需要，并允许产生基因编辑的安乐蜥。这种方法最近已成功应用于其他爬行动物，这表明它可能在广泛的物种中有效，并将广泛地促进对爬行动物基因功能的研究。
