Increasing the effective gene drive homing rate by targeting the haploinsufficient spermatogenesis gene KLHL10
Increasing the effective gene drive homing rate by targeting the haploinsufficient spermatogenesis gene KLHL10
35323Ceili L. Peng, W. Sebastian Kamau, Julien Freeman, et al., bioRxiv, 2026-01-05 12:04:02.
CRISPR-based gene drives represent a powerful new technology for limiting disease transmission and controlling invasive populations. These systems rely on homology-directed repair (HDR) to ‘drive’ a genetic element through a population. However, mammals tend to favor non-homologous end joining (NHEJ), which generates mutations that halt further drive propagation. Here, we describe the experimental characterization of a gene drive system targeting the haploinsufficient spermatogenesis gene KLHL10 in the laboratory mouse. Using a newly designed ‘coding sequence cassette’ we introduce downstream guide RNAs within the gene, ensuring that sperm undergoing NHEJ are selectively removed from the population. As a proof of principle, we demonstrate that targeting KLHL10 with constitutively expressed LbCas12a results in strong selection against frameshift-containing sperm, validating the core purification mechanism required for this drive strategy. Unexpectedly, we also observed that female offspring lacked most frameshift mutations, suggesting a previously unrecognized role for KLHL10 in oogenesis or early embryonic development.
