CRISPR-Mediated Intronic Knockin of Pre-amiRNA Enables Targeted Gene Silencing

We developed an intronic artificial microRNA (IamiRNA) strategy that combines CRISPR/Cas9-mediated knock-in with endogenous miRNA processing for plant gene silencing. By inserting amiRNA precursors into introns of endogenous genes, this approach enables effective and tissue-specific gene silencing without persistent transgene expression, offering a promising tool for functional genomics and crop improvement.