High-frequency gene targeting in Arabidopsis plants expressing the yeast RAD54 gene.
Gene targeting, which is homologous recombination-mediated integration of an extra-chromosomal DNA segment into a chromosomal target sequence, enables the precise disruption or replacement of any gene. Despite its value as a molecular genetic tool, gene targeting remains an inefficient technology in most species. We report that expression of the yeast RAD54 gene, a member of the SWI2/SNF2 chromatin remodeling gene family, enhances gene targeting in Arabidopsis by one to two orders of magnitude, from 10(-4) to 10(-3) in WT plants to 10(-2) to 10(-1). We show that integration events, detected with an assay based on the use of a fluorescent seed marker, are precise and germinally transmitted. These findings suggest that chromatin remodeling is rate-limiting for gene targeting in plants and improves the prospects for using gene targeting for the precise modification of plant genomes.[1]References
- High-frequency gene targeting in Arabidopsis plants expressing the yeast RAD54 gene. Shaked, H., Melamed-Bessudo, C., Levy, A.A. Proc. Natl. Acad. Sci. U.S.A. (2005) [Pubmed]
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