Techniques for Validating CRISPR Changes Using RNA-Sequencing Data.

ABSTRACT: The use of CRISPR to knockdown or knockout genes is a powerful tool for understanding the specific role of a gene in disease development. However, it can cause many unanticipated changes to the transcriptome that are not detected by DNA amplification and Sanger sequencing of the target site. Various RNA-sequencing techniques can be used to identify these changes and effectively gauge the full impact of the CRISPR knockout, thereby providing a means of selecting appropriate clones for further experimentation.

Background/objectives

RNA-seq data from 4 CRISPR knockout experiments were analyzed and techniques developed to both confirm the success of the CRISPR modifications and identify potential issues.

Methods

A broad-based analysis of RNA-sequencing data identified many CRISPR

SUBMITTER: Rathe SK 

PROVIDER: S-EPMC12027141 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature