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An update on CRISPR-Cas12 as a versatile tool in genome editing

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Abstract

Gene editing techniques, which help in modification of any DNA sequence at ease, have revolutionized the world of Genetic engineering. Although there are other gene-editing techniques, CRISPR has emerged as the chief and most preferred tool due to its simplicity and capacity to execute effective gene editing in a wide range of organisms. Although Cas9 has widely been employed for genetic modification over the years, Cas12 systems have lately emerged as a viable option. This review primarily focuses on assessing Cas12-mediated mutagenesis and elucidating the editing efficacy of both Cpf1 (Cas12a) and C2c1 (Cas12b) systems in microbes, plants, and other species. Also, we reviewed several genetic alterations that have been performed with these Cas12 systems to improve editing efficiency. Furthermore, the experimental benefits and applications of Cas12 systems are highlighted in this study.

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Abbreviations

CRISPR:

Clustered regularly interspaced short palindromic repeats

Cas:

CRISPR-associated

crRNA:

CRISPR RNA

TracrRNA:

Trans-acting CRISPR RNA

PAM:

Protospacer adjacent motif

NHEJ:

Non-homologous end joining

HDR:

Homology directed repair

PNSB:

Purple non-sulfur bacteria

DSB:

Double-strand break

TALEN:

Transcription activator-like effector nucleases

ZFN:

Zinc finger nuclease

PEG:

Polyethylene glycol

XCC:

Xanthamonas campestris Pv. campestris

ssODN:

Single stranded oligo donor

PCR:

Polymerase chain reaction

CAR:

Chimeric antigen receptor

FACS:

Fluorescence-activated cell sorting

TIDE:

Tracking of indels by decomposition

CCR5:

CC chemokine receptor 5

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Authors and Affiliations

  1. School of Bio Sciences and Technology, Vellore Institute of Technology, Vellore, 632 014, India

    Raghul Senthilnathan, Ilamathi Ilangovan, Milind Kunale, Nalini Easwaran, Siva Ramamoorthy & Gothandam Kodiveri Muthukaliannan

  2. Center of Excellence in Catalysis for Bioenergy and Renewable Chemicals (CBRC), Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand

    Ashokkumar Veeramuthu

  3. Biorefineries for Biofuels & Bioproducts Laboratory (BBBL), Center for Transdisciplinary Research, Department of Pharmacology, SDC, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, 600077, India

    Ashokkumar Veeramuthu

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  1. Raghul Senthilnathan
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Senthilnathan, R., Ilangovan, I., Kunale, M. et al. An update on CRISPR-Cas12 as a versatile tool in genome editing. Mol Biol Rep 50, 2865–2881 (2023). https://doi.org/10.1007/s11033-023-08239-1

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