绿色荧光蛋白
清脆的
Cas9
表情盒
生物
报告基因
细胞生物学
核定位序列
细胞质
计算生物学
遗传学
基因
基因表达
重组DNA
载体(分子生物学)
作者
Gustavo Aguilar,Oliver Hobert
出处
期刊:PubMed
日期:2024-01-01
卷期号:2024
标识
DOI:10.17912/micropub.biology.000954
摘要
To facilitate cell identification for expression pattern analysis in C. elegans , an SL2::GFP::H2B fluorescent reporter cassette has become a popular and widely used choice to generate nuclear localized reporter alleles by CRISPR/Cas9 genome engineering. When added at the 3' end of a locus of interest, this cassette concentrates GFP into the nucleus and permits the identification of expressing cells, for example with the help of the NeuroPAL tool. However, there are instances in which it is desirable to visualize the complete morphology of a cell that expresses an SL2::GFP::H2B reporter cassette. We describe here a CRISPR/Cas9-engineering strategy to transform an endogenous SL2::GFP::H2B tag into a cytosolic tag by insertion of the self-cleaving T2A tag in between GFP and H2B.
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