1. SKLPT imaging: Efficient in vivo pre-evaluation of genome-editing modules using fluorescent protein with peroxisome targeting signal
- Author
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Ryota Konno, Hiroyuki Tanaka, and Yutaka Kodama
- Subjects
0301 basic medicine ,Green Fluorescent Proteins ,Biophysics ,Computational biology ,Peroxisome localization ,Biochemistry ,Peroxisomal Targeting Signals ,Green fluorescent protein ,03 medical and health sciences ,Marchantia polymorpha ,Genome editing ,Marchantia ,Molecular Biology ,Peroxisomal targeting signal ,Gene Editing ,Transcription activator-like effector nuclease ,biology ,Chemistry ,Cell Biology ,Plants, Genetically Modified ,biology.organism_classification ,Subcellular localization ,Zinc finger nuclease ,Recombinant Proteins ,030104 developmental biology ,Mutagenesis, Site-Directed ,CRISPR-Cas Systems ,Genome, Plant - Abstract
Numerous studies have used genome-editing modules such as CRISPR-Cas9 for site-directed mutagenesis; however, evaluation of the efficiency of these modules remains a time-consuming process. Here, we report the development of SKL-mediated Peroxisome Targeting Imaging (SKLPT imaging), an efficient in vivo pre-evaluation method based on the change in subcellular localization of a fluorescent protein. In this method, frameshifts resulting from successful editing cause the fusion of green fluorescent protein to the peroxisome localization signal Serine-Lysine-Leucine (SKL). Using SKLPT imaging, we pre-evaluated three CRISPR-Cas9 modules in vivo at the single-cell level, and then efficiently mutagenized the liverwort (Marchantia polymorpha) genome using a high-efficiency module.
- Published
- 2018
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