Overcoming the challenges of genome-editing essential genes

This primer manuscript summarizes gene-editing strategies for limiting gene-editing activity with the aim of avoiding biallelic disruption. Mono-allelic editing of cells and embryos is advantageous when modeling dominant genetic disorders or when addressing essential or developmentally important genes, loss of function of which leads to severe phenotypes. Gene-editing reagents, such as the CRISPR-Cas9 system, are very efficient and frequently result in bi-allelic mutation of the selected target site. The article introduces different strategies for restricting editing to a single allele, exploring modifications to both the delivery of the gene-editing reagents and to the enzymes commonly used for gene editing themselves, along with design considerations for both the target sites and the repair template when trying to achieve knockin mutations.