A simple test for the cleavage activity of customized endonucleases in plants.

ABSTRACT:

Background

Although customized endonucleases [transcription activator-like effector nucleases (TALENs) and RNA-guided endonucleases (RGENs)] are known to be effective agents of mutagenesis in various host plants, newly designed endonuclease constructs require some pre-validation with respect to functionality before investing in the creation of stable transgenic plants.

Results

A simple, biolistics-based leaf epidermis transient expression test has been developed, based on reconstituting the translational reading frame of a mutated, non-functional yfp reporter gene. Quantification of mutation efficacy was made possible by co-bombarding the explant with a constitutive mCherry expression cassette, thereby allowing the ratio between the number of red and yellow fluorescing cells

SUBMITTER: Budhagatapalli N 

PROVIDER: S-EPMC4784412 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature