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High-throughput in vitro processing of human primary microRNA by the recombinant microprocessor
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- Authors
- Issue Date
- 2022-03
- Publisher
- Elsevier
- Citation
- STAR Protocols, Vol.3 No.1, p. 101042
- Abstract
- © 2021 The Author(s)We describe a protocol to conduct a high-throughput in vitro processing assay, using 1,881 human primary microRNAs (pri-miRNAs) and recombinant Microprocessor complex, followed by deep sequencing library generation. This comprehensive approach allows the mapping of cleavage sites and the measurement of processing efficiency of a large number of substrates simultaneously. Our protocol is readily modifiable to investigate the effects of chemicals and regulatory proteins. Moreover, cis-acting elements can be examined by replacing the wild-type pri-miRNAs with mutant variants. For complete details on the use and execution of this profile, please refer to Kim et al. (2021).
- ISSN
- 2666-1667
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