Applied BioSciences, Macquarie University, Sydney, NSW, Australia.
Center for Infectious Diseases, Department of Epidemiology, Human Genetics, and Environmental Sciences, School of Public Health, University of Texas Health Science Center, Houston, TX, USA.
Commun Biol. 2024 Sep 12;7(1):1126. doi: 10.1038/s42003-024-06848-5.
CRISPR base editors can introduce point mutations into DNA precisely, and cytosine base editors (CBEs) catalyze C to T transitions. While CBEs have been thoroughly explored in cell culture and organisms such as mice, little is known about DNA base editing in insects. In this study, we evaluated germline editing rates of three different CBEs expressed under actin (ubiquitous) or nanos (germline) promoters utilizing Drosophila melanogaster. The original Rattus norvegicus-derived cytosine deaminase APOBEC1 (rAPO-1) displayed high base editing rates (~99%) with undetectable indel formation. Additionally, we show that base editors can be used for generating male sterility and female lethality. Overall, this study highlights the importance of promoter choice and sex-specific transmission for efficient base editing in flies while providing new insights for future genetic biocontrol designs in insects.
CRISPR 碱基编辑器可以精确地将点突变引入 DNA，而胞嘧啶碱基编辑器（CBEs）则催化 C 到 T 的转换。虽然 CBE 在细胞培养和小鼠等生物中得到了深入研究，但关于昆虫中的 DNA 碱基编辑知之甚少。在这项研究中，我们评估了在黑腹果蝇中利用肌动蛋白（普遍）或纳米（生殖系）启动子表达的三种不同 CBE 的种系编辑率。最初来源于大鼠的胞嘧啶脱氨酶 APOBEC1（rAPO-1）显示出高的碱基编辑率（~99%），并且没有检测到插入缺失的形成。此外，我们还表明，碱基编辑器可用于产生雄性不育和雌性致死。总的来说，这项研究强调了启动子选择和性别特异性传播对于在果蝇中进行高效碱基编辑的重要性，同时为昆虫未来的遗传生物防治设计提供了新的见解。
