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高分辨率原位分析基因驱动的疟蚊 Cas9 种系转录本分布。

High-resolution in situ analysis of Cas9 germline transcript distributions in gene-drive Anopheles mosquitoes.

机构信息

Section of Cell and Developmental Biology, University of California, San Diego, La Jolla, CA 92093, USA.

Tata Institute for Genetics and Society, University of California, San Diego, La Jolla, CA 92093, USA.

出版信息

G3 (Bethesda). 2022 Jan 4;12(1). doi: 10.1093/g3journal/jkab369.

Abstract

Gene drives are programmable genetic elements that can spread beneficial traits into wild populations to aid in vector-borne pathogen control. Two different drives have been developed for population modification of mosquito vectors. The Reckh drive (vasa-Cas9) in Anopheles stephensi displays efficient allelic conversion through males but generates frequent drive-resistant mutant alleles when passed through females. In contrast, the AgNosCd-1 drive (nos-Cas9) in Anopheles gambiae achieves almost complete allelic conversion through both genders. Here, we examined the subcellular localization of RNA transcripts in the mosquito germline. In both transgenic lines, Cas9 is strictly coexpressed with endogenous genes in stem and premeiotic cells of the testes, where both drives display highly efficient conversion. However, we observed distinct colocalization patterns for the two drives in female reproductive tissues. These studies suggest potential determinants underlying efficient drive through the female germline. We also evaluated expression patterns of alternative germline genes for future gene-drive designs.

摘要

基因驱动是可编程的遗传元件,可以将有益性状传播到野生种群中,以帮助控制媒介传播的病原体。已经开发出两种不同的驱动方法来改变蚊子媒介的种群。在斯氏按蚊中,Reckh 驱动(vasa-Cas9)通过雄性表现出高效的等位基因转换,但当通过雌性传递时,会产生频繁的驱动抗性突变等位基因。相比之下,在冈比亚按蚊中,AgNosCd-1 驱动(nos-Cas9)通过两性几乎实现了完全的等位基因转换。在这里,我们检查了蚊子生殖系中的 RNA 转录本的亚细胞定位。在这两种转基因系中,Cas9 与睾丸中的干细胞和减数分裂前细胞中的内源性基因严格共表达,这两种驱动都显示出高效的转换。然而,我们在雌性生殖组织中观察到两种驱动的明显共定位模式。这些研究表明了通过雌性生殖系实现高效驱动的潜在决定因素。我们还评估了替代生殖系基因的表达模式,以用于未来的基因驱动设计。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6049/8728002/9294378d615c/jkab369f1.jpg

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