Synthetic evolution of herbicide resistance using a T7 RNAP-based random DNA base editor.

ABSTRACT: Synthetic directed evolution via localized sequence diversification and the simultaneous application of selection pressure is a promising method for producing new, beneficial alleles that affect traits of interest in diverse species; however, this technique has rarely been applied in plants. Here, we designed, built, and tested a chimeric fusion of T7 RNA Polymerase (RNAP) and deaminase to enable the localized sequence diversification of a target sequence of interest. We tested our T7 RNAP-DNA base editor in Nicotiana benthamiana transient assays to target a transgene expressing GFP under the control of the T7 promoter and observed C-to-T conversions. We then targeted the T7 promoter-driven acetolactate synthase sequence that had been stably integrated in the rice geno

SUBMITTER: Butt H 

PROVIDER: S-EPMC9526444 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature