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通过多重 CRISPR/Cas9 介导的基因失活技术生成透明的食蚊鱼品系。

Generation of a transparent killifish line through multiplex CRISPR/Cas9mediated gene inactivation.

机构信息

Molecular Genetics Lab, Leibniz Institute on Aging - Fritz Lipmann Institute (FLI), Jena, Germany.

Core Facility Imaging, Leibniz Institute on Aging - Fritz Lipmann Institute (FLI), Jena, Germany.

出版信息

Elife. 2023 Feb 23;12:e81549. doi: 10.7554/eLife.81549.

Abstract

Body pigmentation is a limitation for imaging and thus for the performance of longitudinal studies in biomedicine. A possibility to circumvent this obstacle is the employment of pigmentation mutants, which are used in fish species like zebrafish and medaka. To address the basis of aging, the short-lived African killifish has recently been established as a model organism. Despite its short lifespan, shows typical signs of mammalian aging including telomere shortening, accumulation of senescent cells, and loss of regenerative capacity. Here, we report the generation of a transparent line by the simultaneous inactivation of three key loci responsible for pigmentation. We demonstrate that this stable line, named , can serve as a tool for different applications including behavioral experiments and the establishment of a senescence reporter by integration of a fluorophore into the ) locus and microscopy of the resulting line.

摘要

体色素沉着是成像的限制因素,因此也是生物医学中进行纵向研究的限制因素。规避这一障碍的一种可能性是使用色素突变体,这些突变体在斑马鱼和日本青鳉等鱼类物种中被使用。为了研究衰老的基础,寿命较短的非洲脂鲤最近被确立为一种模式生物。尽管其寿命较短,但表现出典型的哺乳动物衰老迹象,包括端粒缩短、衰老细胞积累和再生能力丧失。在这里,我们报告了通过同时失活三个负责色素沉着的关键基因座来产生透明系。我们证明,这条名为 的稳定系可作为不同应用的工具,包括行为实验以及通过将荧光团整合到 )基因座中建立衰老报告基因和对所得系的显微镜观察。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ced7/10010688/3a5b71aa605a/elife-81549-fig1.jpg

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