While homologous recombination-based gene replacement is about to be supplanted by more modern approaches, it is still retaining usefulness for genes that prove to be poor targets for CRISPR/cas-based approaches. Homologous recombination has proven to be relatively robust to minor sequence mismatches between GOI-flanking sequences and the gene replacement constructs, and the faithfulness of recombination events is easily verified by whole-genome sequencing. Moreover, the availability of custom synthetic gene production by numerous service providers should allow for a relatively quick generation of null mutants without the need to introduce additional protein-coding genes beyond the selection markers.
CITATION STYLE
Zirpel, H., & Clos, J. (2019). Gene Replacement by Homologous Recombination. In Methods in Molecular Biology (Vol. 1971, pp. 169–188). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9210-2_8
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