CRISPR/Cas systems versus plant viruses: engineering plant immunity and beyond

Ali, Zahir; Mahfouz, Magdy M.

doi:10.1093/plphys/kiab220

Cited by 14 publications

(10 citation statements)

References 137 publications

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“…Engineering plant immunity to impart plant virus resistance offers great potential for reducing crop losses, increasing plant productivity, and ensuring food security. 26 The CRISPR-Cas9 system has been emerged as a simple, efficient, and precise genome engineering tool for the development of genetically improved crop plants. It has become a method of choice in genome editing experiments in plant research.…”

Section: Discussionmentioning

confidence: 99%

Engineering broad-spectrum resistance to cotton leaf curl disease by CRISPR-Cas9 based multiplex editing in plants

et al. 2021

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Advances in genome editing technologies have tremendous potential to address the limitations of classical resistance breeding. CRISPR-Cas9 based gene editing has been applied successfully in plants to tolerate virus infections. In this study, we successfully tested CRISPR-Cas9 system to counteract cotton leaf curl disease (CLCuD) caused by whitefly transmitted cotton leaf curl viruses (CLCuVs). We also analyzed the ability of CLCuV to escape the Cas9 endonuclease activity. Targeting overlapping genes of most prevalent CLCuVs with three gRNAs resulted in virus interference, as validated by low virus titer. Furthermore, multiplex CRISPR-Cas9 construct simultaneously targeting six genes of CLCuV, was found more effective to interfere with virus proliferation compared to targeting single region individually. Additionally, transgenic N. benthamiana plants expressing multiple gRNAs simultaneously showed enhanced tolerance against CLCuV infection when compared to wild-type plants. T7 Endonuclease-I (T7EI) assay, showing indels in the CLCuV genome, confirmed the occurrence of double strand breaks (DSBs) in DNA at target sequence induced by Cas9 endonuclease. We observed that targeting CLCuV genome at multiple sites simultaneously resulted in better interference, also with inefficient recovery of altered virus molecules. Next, we tested multiplex construct in cotton to interfere CLCuV infection. We found significant decrease in virus accumulation in cotton leaves co-infiltrated with multiplex cassette and virus compared to cotton leaves infiltrated with virus only. The results demonstrate future use of CRISPR-Cas9 system for engineering virus resistance in crops. Moreover, our results also advocate that resistance to mixed virus infections can be engineered using multiplex genome editing.

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Section: Discussionmentioning

confidence: 99%

Engineering broad-spectrum resistance to cotton leaf curl disease by CRISPR-Cas9 based multiplex editing in plants

et al. 2021

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“…By altering five genes relevant to qualities including fruit size, yield, and nutrient content, the Menz team domesticated a wild South American tomato in 2018. It usually takes 8,000 years to perform this process, and “it’s been 1.5 years now” ( El-Mounadi et al., 2020 ; Menz et al, 2020 ; Ali et al., 2021 ; Arya et al., 2021 ; Ali and Mahfouz, 2022 ; Son and Park, 2022 );. Future efforts to get over these restrictions are probably going to expand the experimental freedom and usefulness of the Cas9-CRISPR toolkit.…”

Section: What Benefits Might Crispr/cas-based Gene Editing Offermentioning

confidence: 99%

Plant breeding advancements with “CRISPR-Cas” genome editing technologies will assist future food security

Ahmad

2023

Front. Plant Sci.

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Genome editing techniques are being used to modify plant breeding, which might increase food production sustainably by 2050. A product made feasible by genome editing is becoming better known, because of looser regulation and widespread acceptance. The world’s population and food supply would never have increased proportionally under current farming practices. The development of plants and food production has been greatly impacted by global warming and climate change. Therefore, minimizing these effects is crucial for agricultural production that is sustainable. Crops are becoming more resilient to abiotic stress because of sophisticated agricultural practices and a better understanding of the abiotic stress response mechanism. Both conventional and molecular breeding techniques have been used to create viable crop types both processes are time-consuming. Recently, plant breeders have shown an interest in genome editing approaches for genetic manipulation that use clustered regularly interspaced short palindromic repeats (CRISPR/Cas9). To ensure the security of the food supply in the future, plant kinds with desired traits must be developed. A completely new era in plant breeding has begun because of the revolution in genome editing techniques based on the CRISPR/CRISPR-associated nuclease (Cas9) systems. All plants may effectively target a particular gene or group of loci using Cas9 and single-guide RNA (sgRNA). CRISPR/Cas9 can thereby save time and labor compared to conventional breeding methods. An easy, quick, and efficient method for directly altering the genetic sequences in cells is with the CRISPR and Cas9 systems. The CRISPR-Cas9 system, which was developed from components of the earliest known bacterial immune system, allows for targeted gene breakage and gene editing in a variety of cells/RNA sequences to guide endonuclease cleavage specificity in the CRISPR-Cas9 system. Editing can be directed to practically any genomic site by altering the guide RNA (gRNA) sequence and delivering it to a target cell along with the Cas9 endonuclease. We summarize recent CRISPR/Cas9 plant research findings, investigate potential applications in plant breeding, and make predictions about likely future breakthroughs and approaches to food security through 2050.

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“…In the last few decades, molecular diagnosis methods such as LAMP have evolved. Ali and Mahfouz have distinctly exhibited the application of CRISPR in development of a diagnostic platform for detection of plant viruses, as shown in Figure …”

Section: Recent Developments In Nucleic Acid Based Detectionmentioning

confidence: 99%

Section: Recent Developments In Nucleic Acid Based Detectionmentioning

confidence: 99%

“…Ali and Mahfouz have distinctly exhibited the application of CRISPR in development of a diagnostic platform for detection of plant viruses, as shown in Figure 5. 62 Currently, it is used for the detection of Salmonella in food, bacterial and fungal contamination in food, detection of Listeria monocytogene strains, Verticillium dahliae, Pythium helicoides, Clerotinia sclerotiorum, and diagnosis of plant parasitic nematodes. 63−65 RPA utilizes recombinase enzymes by forming complexes with oligonucleotide primers and pairing the primers with their homologous sequences in duplex DNA.…”

Section: Aptamer Based Approach For Plant Pathogenmentioning

confidence: 99%

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Emerging Extraction and Diagnostic Tools for Detection of Plant Pathogens: Recent Trends, Challenges, and Future Scope

Verma

Shukla

Kulkarni

et al. 2022

ACS Agric. Sci. Technol.

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Plant pathogens are a serious threat to agriculture for long-term viability and cost loss of billions of dollars yearly. Many pathogens have been documented in the literature that infect a variety of crops. Nevertheless, new pathogens emerge and often result in disease outbreaks, leading to million-dollar losses. Currently, several diagnostic approaches with enhanced sensitivity and specificity for the identification of widespread and/or unknown plant pathogens are constantly being developed. Whereas the extensively used approaches for plant pathogen diagnostics are mostly serological and nucleic acid-based assays, many different nucleic acid-based approaches for amplifying target DNA/RNA have also emerged over time. However, these approaches lack precision, specificity, and rapidity, making them unsuitable for on-field analysis. As a result, there is a lot of interest arising in fielddeployable point of care (POC) devices and artificial intelligence (AI)-assisted pathogens' detection accurately at an early stage within a minute. Similarly, development of a cell-lysis and purification-free DNA/RNA extraction process is also crucial for quick sample preparation for molecular diagnosis of plant pathogens at field level. In this review, we have discussed advanced tools that are trending not only to extract nucleic acids but also detect plant pathogens. We have also discussed critical challenges and future perspectives of disease diagnostic tools for plant pathogens' detection. In summary, advanced plant disease diagnostic tools can be helpful for routine monitoring of plant pathogens toward improving crop productivity and yield that can be used for improving the financial status of farmers.

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CRISPR/Cas systems versus plant viruses: engineering plant immunity and beyond

Cited by 14 publications

References 137 publications

Engineering broad-spectrum resistance to cotton leaf curl disease by CRISPR-Cas9 based multiplex editing in plants

Engineering broad-spectrum resistance to cotton leaf curl disease by CRISPR-Cas9 based multiplex editing in plants

Plant breeding advancements with “CRISPR-Cas” genome editing technologies will assist future food security

Emerging Extraction and Diagnostic Tools for Detection of Plant Pathogens: Recent Trends, Challenges, and Future Scope

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