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Author: search.filters.author.Trivedi, Prabodh KumarSubject: search.filters.subject.CRISPR/Cas9

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    Gene pyramiding in transgenic plant development: Approaches and challenges
    (Springer Nature Publishing AG, 2023) Rajput, Ruchika; Naik, Jogindra; Misra, Prashant; Trivedi, Prabodh Kumar; Pandey, Ashutosh
    Climate change leads to frequent alterations in environmental factors with a reciprocal impact on crop productivity. Over the last few decades, various approaches have been used for producing more stress-tolerant and climate-flexible crops. Genetic engineering is one of the approaches used to modify multiple characters or to improve more than one agronomic trait in plants. These instances simultaneously demand simultaneous genetic manipulation of multiple genes, necessitating stacking or pyramiding of multiple genes as compared to single-gene manipulations, and the genetic engineering of plants using multiple genes is technically challenging. In the last two decades, considerable progress has been made with respect to the development and application of the methods for gene pyramiding in transgenic context. The conventional methods of gene stacking include the crossing of individual transgenic plants, co-transformation using multiple plant expression constructs, transformation with single constructs carrying multiple transgenes as well as with the constructs carrying polycistronic transgenes. These methods have been instrumental for gene stacking in several commercialized crops. The tools of targeted genome editing (ZFN, TALEN, and CRISPR) that carry out precise genetic modifications, have opened new avenues in the area of crop biotechnology for defending plants against various stresses. The present review covers the current status of biotechnological techniques used to combat biotic and abiotic stresses in crop plants and describes multiple associated challenges.
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    Emerging tools and paradigm shift of gene editing in cereals, fruits, and horticultural crops for enhancing nutritional value and food security
    (John Wiley & Sons, 2021) Tiwari, Manish; Trivedi, Prabodh Kumar; Pandey, Ashutosh
    Gene editing using sequence‐specific nucleases, particularly CRISPR/Cas ribonucleoprotein, has drawn enormous attention in plant research in recent years. Nearly a decade ago, Cas9 protein was initially discovered for a role in adaptive immunity in bacteria. Owing to vast potential, a large number of reports came out in a short span of time, comprising the identification of Cas protein from different bacterial sources, new Cas9 variants with reduced off‐targets, multiplexing, base editing, prime editing, and RNA manipulation in plants. Studies revealed that CRISPR/Cas‐based gene editing can play a major role in ensuring food security via developing resilient commercial crops with improved yield and nutritional value. Use of the CRISPR/Cas9 system for creating mutation in genes and regulatory regions of promoter generated a number of alleles with variable phenotypes, which can serve as an excellent genetic resource in the breeding program. In this review, we provide a recent overview of state‐of‐art discoveries in the CRISPR/Cas system comprised of new Cas proteins, modifications of existing Cas9, refinements in CRISPR/Cas‐induced gene editing, applications, and outcome emphasizing on major cereals and horticultural crops. We also highlight the current global policy framework for the regulation of gene‐edited crops.