seekRNA Directs Precise Gene Insertion

The study reveals that members of the IS1111 and IS110 insertion sequence (IS) families use a noncoding RNA (NCR)-derived seekRNA to guide target site selection during transposition. This seekRNA, which copurifies with the DEDD transposase, contains sequences complementary to both strands of the target site, essential for transposition.

The seekRNA's configuration differs between the two families, indicating distinct mechanisms of target recognition. The researchers demonstrated the biotechnological potential of reprogramming the seekRNA and donor flank to target different sites, thereby enhancing gene-editing precision.

In experiments, the team showed that seekRNA's role is crucial for the transposition of both the IS element and any cargo it may carry, such as therapeutic genes. The researchers successfully reprogrammed seekRNAs to direct transposition to new target sequences, proving its versatility.

This approach presents a significant advancement in the field of genetic engineering, offering a method to achieve high specificity in gene insertion without introducing unintended mutations. The innovation could pave the way for more precise gene therapy applications, reducing the risk of off-target effects commonly associated with traditional gene-editing technologies.

The study was led by Ruth Hall and Sandro Ataide at the University of Sydney, Australia, and it was published in Nature Communications on June 19, 2024.