CRISPR/Cas9 System to Knockdown MicroRNA In Vitro and In Vivo
Methods in Molecular Biology
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.
Yi, Bin; Larter, Kristina; and Xi, Yaguang, "CRISPR/Cas9 System to Knockdown MicroRNA In Vitro and In Vivo" (2021). School of Medicine Faculty Publications. 85.