Integration of a FT expression cassette into CRISPR/Cas9 construct enables fast generation and easy identification of transgene-free mutants in Arabidopsis

• DOI: 10.1371/journal.pone.0218583
• Publication Date: 2019-09-23T13:28:56Z
• Authors: Yuxin Cheng; Na Zhang; Saddam Hussain; Sajjad Ahmed; Wenting Yang; Shucai Wang
• Keywords: Bolting

The CRISPR/Cas9 genome editing technique has been widely used to generate transgene-free mutants in different plant species. Several methods including fluorescence marker-assisted visual screen of and programmed self-elimination construct have increase the efficiency edited mutant isolation, but overall time length required obtain remained unchanged these methods. We report here a method for fast generation easy identification Arabidopsis. By generating using single FT expression cassette-containing construct, we targeted two sites AITR1 gene. obtained many early bolting plants T1 generation, found that about thirds detectable mutations. then analyzed T2 generations representative lines plants, identified without phenotype, i.e., both lines. Further more, aitr1 homozygous were successful from plants. Taken together, results suggest described enables at mean time,