CRISPR/Cas9 System to Knockdown MicroRNA In Vitro and In Vivo

Document Type

Article

Publication Date

4-1-2021

Publication Title

Methods in Molecular Biology

Abstract

MicroRNAs (miRNAs) are a class of small noncoding single-stranded RNA molecules containing 18–22 nucleotides that play an important role in the regulation of gene expression at the post-transcriptional and translational levels. Loss-of-function studies are the fundamental strategy to examine miRNA function and target genes in cellular and molecular biology. Traditional methods for miRNA loss-of-function studies include miRNA-specific antisense inhibitors, miRNA sponges, and genetic knockout. However, efficiency, specificity, and stability of these methods are not adequate. Our study suggests that CRISPR/Cas9 is an economic, convenient, and innovative strategy with high efficiency, specificity, and stability for the modulation of miRNA expression. Herein, we describe a detailed protocol for knocking out miRNA genes in vitro and in vivo with the CRISPR/Cas9 system.

First Page

133

Last Page

139

PubMed ID

33792878

Volume

2300

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