The Cas9 nuclease has been stably integrated into the host cell genome via genetic engineering methods, enabling stable, long-term expression of Cas9 protein. Cas9 stable cell lines serve as essential tools for CRISPR gene knockout, CRISPR library screening, and functional genomics studies. Ubigene has developed more than 500 Cas9 stable cell lines covering commonly used cell types. Gene knockout can be rapidly achieved by simply transfecting gRNA. Co-transfection of gRNA with donor DNA further enables gene knock-in or precise point mutations.

The Ubigene RKO-Cas9 cell line was generated using a lentiviral method and selected with antibiotics to achieve stable, high-level Cas9 expression. The RKO-CAS9 cell line has undergone functional editing validation, with results demonstrating that introduction of target gene gRNA can produce efficient DNA cleavage and indels, making it suitable for gene knockout, gRNA efficiency validation, or high-throughput CRISPR library screening in RKO cells.