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Identification of a Novel Cleavage Site and Confirmation of the Effectiveness of NgAgo Gene Editing on RNA Targets

Mol Biotechnol. 2021-07; 
Jiayao Qu, Yali Xie, Zhaoyi Guo, Xiangting Liu, Jing Jiang, Ting Chen, Kai Li, Zheng Hu, Dixian Luo
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Nucleic Acid Purification & Analysis … studies were manufactured by GenScript (Nanjing, … out using the primers in the kit and the synthetic primers T7-GFP-PCR-F or T7-HCV-PCF-F (Table S1). The amplicons were separated using agarose gel electrophoresis, and the PCR products were extracted and purified using … Get A Quote

摘要

Clusters of regularly interspaced short palindromic repeats (CRISPR)/Cas systems have a powerful ability to edit DNA and RNA targets. However, the need for a specific recognition site, protospacer adjacent motif (PAM), of the CRISPR/Cas system limits its application in gene editing. Some Argonaute (Ago) proteins have endonuclease functions under the guidance of 5' phosphorylated or hydroxylated guide DNA (gDNA). The NgAgo protein might perform RNA gene editing at 37 °C, suggesting its application in mammalian cells; however, its mechanisms are unclear. In the present study, the target of NgAgo in RNA was confirmed in vitro and in vivo. Then, an in vitro RNA cleavage system was designed and the cleavage site w... More

关键词

Ago, Argonaute, Cleavage site, NgAgo, RNA editing, gDNA