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Recombinant tag-free Leptotrichia buccalis LbuCas13a was expressed in E. coli cells.
CS13A-E322U-10 |
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概述:
Cas13a is an RNA-guided RNA endonuclease that belongs to the class 2 type VI CRISPR-Cas system (1). Cas13a possesses both RNA recognition and cleavage activities and cleaves RNA through conserved basic residues within its two HEPN domains (2). Interestingly, activated Cas13a induces collateral cleavage of nearby non-targeted RNAs in a nonspecific manner (3). Engineered LbuCas13a shows stability and altered cleavage activity in vitro and possesses the capability of robust exon skipping in human cells (4).
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参考文献:
1. East-Seletsky A, et al: Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016, 538:270–3.
2. Abudayyeh O, et al: C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 2016, 353:5573.
3. Patchsung M, et al: Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA. Nat Biomed Eng. 2020. https://doi.org/10.1038/s41551-020-00603-x.
4. Charles E, et al: Engineering improved Cas13 effectors for targeted post-transcriptional regulation of gene expression. bioRxiv. doi: https://doi.org/10.1101/2021.05.26.445687.
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