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A CRISPR-Cas and Tat Peptide with Fluorescent RNA Aptamer System for Signal Amplification in RNA Imaging
oleh: Heng Tang, Junran Peng, Xin Jiang, Shuang Peng, Fang Wang, Xiaocheng Weng, Xiang Zhou
Format: | Article |
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Diterbitkan: | MDPI AG 2023-02-01 |
Deskripsi
We reported on an efficient RNA imaging strategy based on a CRISPR-Cas and Tat peptide with a fluorescent RNA aptamer (TRAP-tag). Using modified CRISPR-Cas RNA hairpin binding proteins fused with a Tat peptide array that recruits modified RNA aptamers, this simple and sensitive strategy is capable of visualizing endogenous RNA in cells with high precision and efficiency. In addition, the modular design of the CRISPR-TRAP-tag facilitates the substitution of sgRNAs, RNA hairpin binding proteins, and aptamers in order to optimize imaging quality and live cell affinity. With CRISPR-TRAP-tag, exogenous <i>GCN4</i>, endogenous mRNA <i>MUC4</i>, and lncRNA <i>SatIII</i> were distinctly visualized in single live cells.