Small-Molecule-Mediated Split-Aptamer Assembly for Inducible CRISPR-dCas9 Transcription Activation

Autor:	Xiao-Han Liu, Bang-Rui Li, Zhan-Ming Ying, Li-Juan Tang, Fenglin Wang, Jian-Hui Jiang
Rok vydání:	2022
Předmět:	Transcriptional Activation S-Adenosylmethionine Gene Expression Regulation Molecular Medicine Clustered Regularly Interspaced Short Palindromic Repeats General Medicine CRISPR-Cas Systems Biochemistry RNA Guide Kinetoplastida
Zdroj:	ACS Chemical Biology. 17:1769-1777
ISSN:	1554-8937 1554-8929
DOI:	10.1021/acschembio.2c00101
Popis:	Inducible CRISPR-dCas9 transcription system has become a powerful tool for transcription regulation and sensing. Here, we develop a new concept of small-molecule-mediated split-aptamer assembly for inducible CRISPR-dCas9 transcription activation, allowing quantitative detection and imaging of S-adenosyl methionine (SAM) in live cells. This inducible transcription system is designed by integrating one fragment of a split SAM aptamer to guide RNA (gRNA) and the other to MS2 arrays. SAM-mediated reassembly of the split fragments recruits an MCP-fused transcription activator to the gRNA-dCas9 complex, activating the expression of a near-infrared fluorescent protein for imaging. We demonstrate that this inducible transcription system achieves quantitative detection of SAM with high sensitivity in live cells. Our system shows that methionine adenosyltransferase 1A (MAT1A) and MAT2A can both catalyze SAM production in live cells and the SAM levels in cancer cells can be increased via upregulation of MAT1A mRNA by epigenetic inhibitors. This split-aptamer assembly strategy could afford a new approach for controlling the CRISPR-dCas9 system, enabling conditional transcription regulation in response to endogenous metabolites in live cells.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::20e3eab2e4165f6803bae900cde1dd09 https://doi.org/10.1021/acschembio.2c00101 Zobrazit plný text záznamu