期刊
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
卷 143, 期 47, 页码 19834-19843出版社
AMER CHEMICAL SOC
DOI: 10.1021/jacs.1c09041
关键词
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资金
- National Natural Science Foundation of China [21977122, 21708054]
- Innovative Research Team in University of Ministry of Education of China [IRT_17R111]
- National Key R&D Program of China [2020YFA0211200]
This study introduces a self-assembly-induced RNA circuit that can establish regulatory connections between endogenous genes, successfully programming the cellular phenotype of E. coli and mammalian cells.
Construction of synthetic circuits that can artificially establish endogenous gene connections is essential to introduce new phenotypes for cellular behaviors. Given the diversity of endogenous genes, it lacks a general and easy-to-design toolbox to manipulate the genetic network. Here we present a type of self-assembly-induced RNA circuit that can directly build regulatory connections between endogenous genes. Inspired from the natural assembling process of guide RNA in the CRISPR/Cas9 complex, this design employs an independent trigger RNA strand to induce the formation of a ternary guide RNA assembly for functional control of CRISPR/Cas9. With this general principle, expressional regulations of endogenous genes can be controlled by totally independent endogenous small RNAs and mRNAs in E. coli via activatable CRISPR/Cas9 function. Moreover, the cellular phenotype of E. coli is successfully programmed with introduction of new gene connections. In addition, the functionality of this design is also verified in the mammalian system. This self-assembly-based RNA circuit exhibits a great flexibility and simplicity of design and provides a unique approach to build endogenous gene connections, which paves a broad way toward manipulation of cellular genetic networks.
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