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A review of CRISPR associated genome engineering: application, advances and future prospects of genome targeting tool for crop improvement

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Abstract

The Cas9 nuclease initiates double-stranded breaks at the target position in DNA, which are repaired by the intracellular restoration pathways to eliminate or insert pieces of DNA. CRISPR-Cas9 is proficient and cost-effective since cutting is guided by a piece of RNA instead of protein. Emphasis on this technology, in contrast with two recognized genome editing platforms (i.e., ZFNs and TALENs), is provided. This review evaluates the benefits of chemically synthesized gRNAs as well as the integration of chemical amendments to improve gene editing efficiencies. CRISPR is an indispensable means in biological investigations and is now as well transforming varied fields of biotechnology and agriculture. Recent advancement in targetable epigenomic-editing tools allows researchers to dispense direct functional and transcriptional significance to locus-explicit chromatin adjustments encompassing gene regulation and editing. An account of diverse sgRNA design tools is provided, principally on their target competence prediction model, off-target recognition algorithm, and generation of instructive annotations. The modern systems that have been utilized to deliver CRISPR-Cas9 in vivo and in vitro for crop improvement viz. nutritional enhancement, production of drought-tolerant and disease-resistant plants, are also highlighted. The conclusion is focused on upcoming directions, biosafety concerns, and expansive prospects of CRISPR technologies.

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This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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NKS wrote the initial draft of the manuscript; SA and PS helped with the literature review; SA prepared the manuscript; SA and PS contributed to the several revisions of the manuscript.

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Correspondence to Nand K. Singh.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Afzal, S., Sirohi, P. & Singh, N.K. A review of CRISPR associated genome engineering: application, advances and future prospects of genome targeting tool for crop improvement. Biotechnol Lett 42, 1611–1632 (2020). https://doi.org/10.1007/s10529-020-02950-w

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  • DOI: https://doi.org/10.1007/s10529-020-02950-w

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