CRISPR-Mediated Engineering across the Central Dogma in Plant Biology for Basic Research and Crop Improvement

Pramanik, Dibyajyoti; Shelake, Rahul Mahadev; Kim, Mi Jung; Kim, Jae‐Yean

doi:10.1016/j.molp.2020.11.002

Cited by 75 publications

(45 citation statements)

References 258 publications

(276 reference statements)

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“…Diverse Cas orthologs and variants are available with different protospacer adjacent motif (PAM) requirements, gRNA scaffolds, temperature tolerances, and editing efficiencies. All these tools enable the broad application of CRISPR technology to plant breeding and fundamental research ( Chen et al., 2019 ; Pramanik et al., 2020 ; Veillet et al., 2020 ).…”

Section: Introductionmentioning

confidence: 99%

CRISPR ribonucleoprotein-mediated genetic engineering in plants

Zhang

Iaffaldano

2021

Plant Communications

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CRISPR-derived biotechnologies have revolutionized the genetic engineering field and have been widely applied in basic plant research and crop improvement. Commonly used Agrobacterium - or particle bombardment-mediated transformation approaches for the delivery of plasmid-encoded CRISPR reagents can result in the integration of exogenous recombinant DNA and potential off-target mutagenesis. Editing efficiency is also highly dependent on the design of the expression cassette and its genomic insertion site. Genetic engineering using CRISPR ribonucleoproteins (RNPs) has become an attractive approach with many advantages: DNA/transgene-free editing, minimal off-target effects, and reduced toxicity due to the rapid degradation of RNPs and the ability to titrate their dosage while maintaining high editing efficiency. Although RNP-mediated genetic engineering has been demonstrated in many plant species, its editing efficiency remains modest, and its application in many species is limited by difficulties in plant regeneration and selection. In this review, we summarize current developments and challenges in RNP-mediated genetic engineering of plants and provide future research directions to broaden the use of this technology.

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Section: Introductionmentioning

confidence: 99%

CRISPR ribonucleoprotein-mediated genetic engineering in plants

Zhang

Iaffaldano

2021

Plant Communications

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“…Now, with the advent of the CRISPR/Cas, a revolutionary genome/gene editing tool, the regulatory barrier is expected to get weaken in coming years [274][275][276] as the regulatory agencies of several countries such as USA, Canada, China, etc., have considered them as mutants [276]. In addition, the technique CRISPR/Cas can more favorably modified and used as several variants of Cas enzymes are now available [277]. In the present scenario, CRIPSR/Cas is considered as one of the best tools for editing the traits in crop(s) species.…”

Section: Public Perception For the Metabolic Engineered Plantsmentioning

confidence: 99%

Metabolomics Intervention Towards Better Understanding of Plant Traits

Sharma

Gupta

Priscilla

et al. 2021

Cells

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The majority of the most economically important plant and crop species are enriched with the availability of high-quality reference genome sequences forming the basis of gene discovery which control the important biochemical pathways. The transcriptomics and proteomics resources have also been made available for many of these plant species that intensify the understanding at expression levels. However, still we lack integrated studies spanning genomics–transcriptomics–proteomics, connected to metabolomics, the most complicated phase in phenotype expression. Nevertheless, for the past few decades, emphasis has been more on metabolome which plays a crucial role in defining the phenotype (trait) during crop improvement. The emergence of modern high throughput metabolome analyzing platforms have accelerated the discovery of a wide variety of biochemical types of metabolites and new pathways, also helped in improving the understanding of known existing pathways. Pinpointing the causal gene(s) and elucidation of metabolic pathways are very important for development of improved lines with high precision in crop breeding. Along with other -omics sciences, metabolomics studies have helped in characterization and annotation of a new gene(s) function. Hereby, we summarize several areas in the field of crop development where metabolomics studies have made its remarkable impact. We also assess the recent research on metabolomics, together with other omics, contributing toward genetic engineering to target traits and key pathway(s).

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“…During the error-prone DNA repair process, mutations can be generated. The CRISPR/Cas technology has been successfully applied for generating designer crop varieties, including pathogen-resistant crops [17,20]. In recent times, either the pathogen genome or the host plant genes have been targeted by CRISPR/Cas to gain a disease-resistant phenotype.…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas9-Mediated Generation of Pathogen-Resistant Tomato against Tomato Yellow Leaf Curl Virus and Powdery Mildew

Pramanik

Shelake

Park

et al. 2021

IJMS

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Tomato is one of the major vegetable crops consumed worldwide. Tomato yellow leaf curl virus (TYLCV) and fungal Oidium sp. are devastating pathogens causing yellow leaf curl disease and powdery mildew. Such viral and fungal pathogens reduce tomato crop yields and cause substantial economic losses every year. Several commercial tomato varieties include Ty-5 (SlPelo) and Mildew resistance locus o 1 (SlMlo1) locus that carries the susceptibility (S-gene) factors for TYLCV and powdery mildew, respectively. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) is a valuable genome editing tool to develop disease-resistant crop varieties. In this regard, targeting susceptibility factors encoded by the host plant genome instead of the viral genome is a promising approach to achieve pathogen resistance without the need for stable inheritance of CRISPR components. In this study, the CRISPR/Cas9 system was employed to target the SlPelo and SlMlo1 for trait introgression in elite tomato cultivar BN-86 to confer host-mediated immunity against pathogens. SlPelo-knockout lines were successfully generated, carrying the biallelic indel mutations. The pathogen resistance assays in SlPelo mutant lines confirmed the suppressed accumulation of TYLCV and restricted the spread to non-inoculated plant parts. Generated knockout lines for the SlMlo1 showed complete resistance to powdery mildew fungus. Overall, our results demonstrate the efficiency of the CRISPR/Cas9 system to introduce targeted mutagenesis for the rapid development of pathogen-resistant varieties in tomato.

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CRISPR-Mediated Engineering across the Central Dogma in Plant Biology for Basic Research and Crop Improvement

Cited by 75 publications

References 258 publications

CRISPR ribonucleoprotein-mediated genetic engineering in plants

CRISPR ribonucleoprotein-mediated genetic engineering in plants

Metabolomics Intervention Towards Better Understanding of Plant Traits

CRISPR/Cas9-Mediated Generation of Pathogen-Resistant Tomato against Tomato Yellow Leaf Curl Virus and Powdery Mildew

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