Sharpening gene editing toolbox in Arabidopsis for plants
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REVIEW ARTICLE
Sharpening gene editing toolbox in Arabidopsis for plants Sagar S. Arya1 • Binod K. Mahto1 • Thakku R. Ramkumar2 • Sangram K. Lenka1 Received: 2 April 2020 / Accepted: 8 September 2020 Society for Plant Biochemistry and Biotechnology 2020
Abstract Arabidopsis thaliana is considered as an indispensable model system across various disciplines of modern plant biology. The short life cycle, well developed classical genetics, small genome, and a plethora of distinguishable phenotypic features make it the most preferred plant system for researchers globally. Owing to these advantages, it is highly explored to study plant-pathogen interaction, comparative omics, evolutionary plasticity, epigenetics, and recently being used for development of precision genome editing tools for plants. Clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated (Cas) protein system, discovered as a part of bacterial immune system is widely used as a molecular tool for targeted genome engineering. This tool superseded two well-established genome editing platforms, zinc finger nucleases and transcription activator-like effector nucleases. Here, we have discussed CRISPR/Cas based gene editing toolbox designed, sharpened and used in A. thaliana. This review focuses on pioneering innovations in CRISPR/Cas technology and subsequent application of these tools in A. thaliana which is leading to wider application of CRISPR toolbox for basic and translational research in other plants. Keywords Arabidopsis thaliana Genome editing CRISPR/Cas base editing CRISPR-TSKO Abbreviations CRISPR/Cas TALENs RNPs ZFNs DSB NHEJ HDR MMEJ BE CBEs ABEs ADARs Indels
CRSIPR-TSKO Clustered regularly interspaced short palindromic repeats/Cas associated. Transcription activator-like effector nucleases Ribonucleoproteins Zinc finger nucleases Double-stranded break Non-homologous end joining Homology-directed repair Microhomology mediated end joining pathway Base editing Cytidine base editors Adenine base editors Adenosine deaminases act on RNA Insertion-deletion mutations
& Sangram K. Lenka [email protected] 1
TERI-Deakin NanoBiotechnology Centre, The Energy and Resources Institute, New Delhi 110003, India
2
Agronomy Department, IFAS, University of Florida, Gainesville, FL 32611, USA
Clustered regularly interspaced short palindromic repeats-tissue-specific knockout
Introduction Arabidopsis thaliana (a crucifer) is the most widely studied model system in plant biology. It has a short life cycle with miniature morphology which can be easily confined to the laboratory environment, well developed classical genetics, small genome with remarkably little dispersed DNA repetition, and possesses diverse morpho-physiological and metabolic phenotypes (Pruitt and Meyerowitz 1986). Moreover, the availability of extensive genomics information of A. thaliana enables researchers to investigate the fundamentals of plant molecular biology. Thus, owing to the in-depth biological understanding, A. thaliana is widely used as a model
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