Development of CRISPR/Cas9 mediated virus resistance in agriculturally important crops

Khatodia, Surender; Bhatotia, Kirti; Tuteja, Narendra

doi:10.1080/21655979.2017.1297347

Cited by 43 publications

(25 citation statements)

References 24 publications

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“…For example, an SaCas9/gRNA system was programed to speci cally target HIV-1 provirus and suppress HIV-1 infection in Jurkat C11 cells [15]. CRISPR/Cas9 was also employed to target plant viruses, which provided a more versatile viral resistance in important crops [16]. Class II CRISPR systems include Cas13a other than cas9 [17].…”

Section: Introductionmentioning

confidence: 99%

A programmable CRISPR/Cas9-based phage defense system for Escherichia coli BL21(DE3)

Liu

Zhao

et al. 2020

Preprint

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Abstract Escherichia coli BL21 is arguably the most popular host for industrial production of proteins, and industrial fermentations are often plagued by phage infections. The CRISPR/Cas system is guided by a gRNA to cleave a specific DNA cassette, which can be developed into a highly efficient programable phage defense system. In this work, we constructed a CRISPR/Cas system targeting multiple positions on the genome of T7 phage and found that the system increased the BL21’s defense ability against phage infection. Furthermore, the targeted loci on phage genome played a critical role. For better control of expression of CRISPR/Cas9, various modes were tested, and the OD of the optimized strain BL21(pT7cas9, pT7-3gRNA, prfp) after 4 hours of phage infection was significantly improved, reaching 2.0, which was similar to the control culture without phage infection. Although at later time points, the defensive ability of CRISPR/Cas9 systems were not as obvious as that at early time points. The viable cell count of the engineered strain in the presence of phage was only one order of magnitude lower than that of the strain with no infection, which further demonstrated the effectiveness of the CRISPR/Cas9 phage defense system. Finally, the engineered BL21 strain under phage attack expressed RFP protein at about 60% of the un-infected control, which was significantly higher than the parent BL21. In this work, we successfully constructed a programable CRISPR/Cas9 system to increase the ability of E. coli BL21’s to defend against phage infection, and created a resistant protein expression host. This work provides a simple and feasible strategy for protecting industrial E. coli strains against phage infection.

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Section: Introductionmentioning

confidence: 99%

A programmable CRISPR/Cas9-based phage defense system for Escherichia coli BL21(DE3)

Liu

Zhao

et al. 2020

Preprint

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“…Novel therapeutic strategies are urgently required to treat such infectious viruses. Since its discovery, the CRISPR/Cas9 system has been developed into a multifunctional genetic modification tool, which can now be used to perform gene deletion, gene insertion, genome‐wide screening, gene therapy and genetic breeding (Khatodia et al , ; Ledford, ; Zhang et al , ; Gebre et al , ). In previous studies, we demonstrated that the system can effectively edit the BmNPV genome by targeting the ie‐1 gene, thereby inhibiting the viral DNA replication in vivo and in vitro (Dong et al , ; Dong et al , ).…”

Section: Discussionmentioning

confidence: 99%

CRISPR/Cas9‐mediated disruption of the immediate early‐0 and 2 as a therapeutic approach to Bombyx mori nucleopolyhedrovirus in transgenic silkworm

Dong

et al. 2018

Insect Molecular Biology

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The CRISPR/Cas9 system is a powerful tool for the treatment of infectious diseases. In our previous study, we knocked out the Bombyx mori nucleopolyhedrovirus (BmNPV) key genes and BmNPV-dependent host factor to generate transgenic antiviral strains. To further expand the range of target genes for BmNPV and more effectively prevent and control pathogenic infections, we performed gene editing and antiviral analysis by constructing a target-directed baculovirus early transcriptional activator immediate early-0 (ie-0) and 2 (ie-2) transgenic silkworm line. We hybridized it with Cas9 transgenic line to produce a double-positive transgenic Cas9(+)/sgIE0-sgIE2(+) line that could activate the CRISPR gene editing system. We first demonstrated that the system is capable of efficiently editing target genes and resulting in fragment deletions in the BmNPV genome. Survival rate of the transgenic Cas9(+)/sgIE0-sgIE2(+) line reached 65% after inoculation with 1 × 10 occlusion bodies/larva. Molecular analysis showed that BmNPV DNA replication and viral gene expression level in the transgenic Cas9(+)/sgIE0-sgIE2(+) line were significantly inhibited compared with the control Cas9(-)/sgIE0-sgIE2(-) line. These results indicated that IE-0 and IE-2, as baculovirus early transcriptional activators, can be used as target sites for gene therapy and that multigene editing could expand the range of target sites for research to create silkworm resistance breeds.

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“…The CRISPR/Cas9 tools for key genome editing are currently applied for reporting virus resistance in the plants [23]. CRISPR-Cas9 is an RNA directed DNA endonuclease organization, which comprises of Cas9 nuclease showing limited targeting bases through protospacer end-to-end motif complexed by simply customizable sole controller RNA targeting about 20-bp genomic arrangement.…”

Section: Global Perspective On Pharmaceuticals Bioengineered Toolsmentioning

confidence: 99%

Bioengineering tools for the production of pharmaceuticals: current perspective and future outlook

2019

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The bioengineering tools have significant advantages through less time-consuming and utilized as a promising stage for the production of pharmaceutical bioproducts under the single platform. This review highlighted the advantages and current improvement in the plant, animal and microbial bioengineering tools and outlines feasible approaches by biological and process's bioengineering levels for advancing the economic feasibility of pharmaceutical's production. The critical analysis results revealed that system biology and synthetic biology along with advanced bioengineering tools like transcriptome, proteome, metabolome and nano bioengineering tools have shown a promising impact on the development of pharmaceutical's bioproducts. Tools to overcome and resolve the accompanying encounters of pharmaceutical's production that include nano bioengineering tools are also discussed. As a summary and prospect, it also gives new insight into the challenges and possible breakthrough of the development of pharmaceutical's bioproducts through bioengineering tools.

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Development of CRISPR/Cas9 mediated virus resistance in agriculturally important crops

Cited by 43 publications

References 24 publications

A programmable CRISPR/Cas9-based phage defense system for Escherichia coli BL21(DE3)

A programmable CRISPR/Cas9-based phage defense system for Escherichia coli BL21(DE3)

CRISPR/Cas9‐mediated disruption of the immediate early‐0 and 2 as a therapeutic approach to Bombyx mori nucleopolyhedrovirus in transgenic silkworm

Bioengineering tools for the production of pharmaceuticals: current perspective and future outlook

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