Gene targeting in Arabidopsis via an all-in-one strategy that uses a translational enhancer to aid Cas9 expression.

Gene targeting, i.e. precise sequence knock-in or replacement, is the ultimate goal of precision genome engineering. Although homologous recombination (HR)-mediated gene targeting is widely used in many organisms, including mammals (Thomas and Capecchi, 1987) (Chandrasegaran and Carroll, 2016), the low frequencies of HR in higher plants limit its utilization for gene targeting (Fauser et al., 2012). We recently described a sequential transformation approach for efficient (5.3~9.1%) HR-mediated gene targeting in Arabidopsis.