通过 CRISPR/Cas9 介导的 LexA 基因敲入,靶向删除黑腹蝇中的嗅觉受体。

IF 1.8 4区 医学 Q3 GENETICS & HEREDITY
Runqi Zhang, Renny Ng, Shiuan-Tze Wu, Chih-Ying Su
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引用次数: 0

摘要

对黑腹果蝇嗅觉的研究极大地受益于嗅觉受体突变体系和 GAL4 报告基因系等遗传试剂。CRISPR/Cas9 基因编辑系统被越来越多地用于创建无效受体突变体或用 GAL4 报告基因替换编码区。为了进一步扩展这一操纵蝇类嗅觉受体神经元(ORNs)的工具包,我们利用 CRISPR/Cas9 基因敲入 LexA 驱动因子,产生了 11 种不同嗅觉受体的空等位基因,其中包括迄今为止还没有敲入突变体的多种受体系。这些目标神经元类型代表了来自所有四种形态感觉器类别的多种触角 ORN。此外,我们还证实了它们的功能缺失表型,评估了受体的单倍体效率,并评估了 LexA 基因敲入驱动的特异性。这些受体突变品系已存放在布卢明顿果蝇种群中心,供更广泛的科学界使用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Targeted deletion of olfactory receptors in D. melanogaster via CRISPR/Cas9-mediated LexA knock-in.

The study of olfaction in Drosophila melanogaster has greatly benefited from genetic reagents such as olfactory receptor mutant lines and GAL4 reporter lines. The CRISPR/Cas9 gene-editing system has been increasingly used to create null receptor mutants or replace coding regions with GAL4 reporters. To further expand this toolkit for manipulating fly olfactory receptor neurons (ORNs), we generated null alleles for 11 different olfactory receptors by using CRISPR/Cas9 to knock in LexA drivers, including multiple lines for receptors which have thus far lacked knock-in mutants. The targeted neuronal types represent a broad range of antennal ORNs from all four morphological sensillum classes. Additionally, we confirmed their loss-of-function phenotypes, assessed receptor haploinsufficiency, and evaluated the specificity of the LexA knock-in drivers. These receptor mutant lines have been deposited at the Bloomington Drosophila Stock Center for use by the broader scientific community.

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来源期刊
Journal of neurogenetics
Journal of neurogenetics 医学-神经科学
CiteScore
4.40
自引率
0.00%
发文量
13
审稿时长
>12 weeks
期刊介绍: The Journal is appropriate for papers on behavioral, biochemical, or cellular aspects of neural function, plasticity, aging or disease. In addition to analyses in the traditional genetic-model organisms, C. elegans, Drosophila, mouse and the zebrafish, the Journal encourages submission of neurogenetic investigations performed in organisms not easily amenable to experimental genetics. Such investigations might, for instance, describe behavioral differences deriving from genetic variation within a species, or report human disease studies that provide exceptional insights into biological mechanisms
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