Comparing engineered nuclear-localized reporter cassettes.
Comparing engineered nuclear-localized reporter cassettes.
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DOI：
10.17912/micropub.biology.001014
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发表时间：
2023
影响因子：
--
通讯作者：
Hobert, Oliver
中科院分区：
文献类型：
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作者：
Sun, HaoSheng;Beets, Isabel;Schafer, William;Hobert, Oliver
最近的单细胞转录组分析揭示了秀丽隐杆线虫神经系统中神经肽编码基因表达的巨大广度和特异性。为了分析神经肽基因表达的动态，以及剖析其表达受控制的调控机制，报告基因仍然是一个重要的工具。利用CRISPR/Cas9基因组工程技术，我们生成了6个不同神经肽编码基因的报告等位基因（3个flp基因，1个nlp基因和2个胰岛素基因）。我们发现不同的报告者磁带导致不同水平的报告者表达，并建议使用SL2::GFP::H2B或GFP::H2B::SL2磁带。
Recent single-cell transcriptome analysis has revealed a tremendous breadth and specificity of neuropeptide-encoding gene expression in the nervous system of C. elegans. To analyze the dynamics of neuropeptide gene expression, as well as to dissect the regulatory mechanism by which their expression is controlled, reporter genes remain an important tool. Using CRISPR/Cas9 genome-engineering, we generate here reporter alleles for 6 different neuropeptide encoding genes (3 flp genes, 1 nlp and 2 insulin genes). We find that different reporter cassettes result in different levels of reporter expression and recommend usage of an SL2::GFP::H2B or GFP::H2B::SL2 cassette.