The Experts below are selected from a list of 213 Experts worldwide ranked by ideXlab platform
Magdy M Mahfouz - One of the best experts on this subject based on the ideXlab platform.
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crispr cas13d mediates robust rna Virus Interference in plants
Genome Biology, 2019Co-Authors: Ahmed Mahas, Rashid Aman, Magdy M MahfouzAbstract:CRISPR-Cas systems endow bacterial and archaeal species with adaptive immunity mechanisms to fend off invading phages and foreign genetic elements. CRISPR-Cas9 has been harnessed to confer Virus Interference against DNA Viruses in eukaryotes, including plants. In addition, CRISPR-Cas13 systems have been used to target RNA Viruses and the transcriptome in mammalian and plant cells. Recently, CRISPR-Cas13a has been shown to confer modest Interference against RNA Viruses. Here, we characterized a set of different Cas13 variants to identify those with the most efficient, robust, and specific Interference activities against RNA Viruses in planta using Nicotiana benthamiana. Our data show that LwaCas13a, PspCas13b, and CasRx variants mediate high Interference activities against RNA Viruses in transient assays. Moreover, CasRx mediated robust Interference in both transient and stable overexpression assays when compared to the other variants tested. CasRx targets either one Virus alone or two RNA Viruses simultaneously, with robust Interference efficiencies. In addition, CasRx exhibits strong specificity against the target Virus and does not exhibit collateral activity in planta. Our data establish CasRx as the most robust Cas13 variant for RNA Virus Interference applications in planta and demonstrate its suitability for studying key questions relating to Virus biology.
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engineering rna Virus Interference via the crispr cas13 machinery in arabidopsis
Viruses, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Magdy M MahfouzAbstract:Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) systems are key immune mechanisms helping prokaryotic species fend off RNA and DNA Viruses. CRISPR/Cas9 has broad applications in basic research and biotechnology and has been widely used across eukaryotic species for genome engineering and functional analysis of genes. The recently developed CRISPR/Cas13 systems target RNA rather than DNA and thus offer new potential for transcriptome engineering and combatting RNA Viruses. Here, we used CRISPR/LshCas13a to stably engineer Arabidopsis thaliana for Interference against the RNA genome of Turnip mosaic Virus (TuMV). Our data demonstrate that CRISPR RNAs (crRNAs) guiding Cas13a to the sequences encoding helper component proteinase silencing suppressor (HC-Pro) or GFP target 2 (GFP-T2) provide better Interference compared to crRNAs targeting other regions of the TuMV RNA genome. This work demonstrates the exciting potential of CRISPR/Cas13 to be used as an antiviral strategy to obstruct RNA Viruses, and encourages the search for more robust and effective Cas13 variants or CRISPR systems that can target RNA.
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Engineering resistance against Tomato yellow leaf curl Virus via the CRISPR/Cas9 system in tomato
Plant signaling & behavior, 2018Co-Authors: Manal Tashkandi, Zahir Ali, Fatimah R Aljedaani, Ashwag Shami, Magdy M MahfouzAbstract:CRISPR/Cas systems confer molecular immunity against phages and conjugative plasmids in prokaryotes. Recently, CRISPR/Cas9 systems have been used to confer Interference against eukaryotic Viruses. Here, we engineered Nicotiana benthamiana and tomato (Solanum lycopersicum) plants with the CRISPR/Cas9 system to confer immunity against the Tomato yellow leaf curl Virus (TYLCV). Targeting the TYLCV genome with Cas9-single guide RNA at the sequences encoding the coat protein (CP) or replicase (Rep) resulted in efficient Virus Interference, as evidenced by low accumulation of the TYLCV DNA genome in the transgenic plants. The CRISPR/Cas9-based immunity remained active across multiple generations in the N. benthamiana and tomato plants. Together, our results confirmed the efficiency of the CRISPR/Cas9 system for stable engineering of TYLCV resistance in N. benthamiana and tomato, and opens the possibilities of engineering Virus resistance against single and multiple infectious Viruses in other crops.
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rna Virus Interference via crispr cas13a system in plants
Genome Biology, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Muhammad Zuhaib Khan, Shouwei Ding, Magdy M MahfouzAbstract:CRISPR/Cas systems confer immunity against invading nucleic acids and phages in bacteria and archaea. CRISPR/Cas13a (known previously as C2c2) is a class 2 type VI-A ribonuclease capable of targeting and cleaving single-stranded RNA (ssRNA) molecules of the phage genome. Here, we employ CRISPR/Cas13a to engineer Interference with an RNA Virus, Turnip Mosaic Virus (TuMV), in plants. CRISPR/Cas13a produces Interference against green fluorescent protein (GFP)-expressing TuMV in transient assays and stable overexpression lines of Nicotiana benthamiana. CRISPR RNA (crRNAs) targeting the HC-Pro and GFP sequences exhibit better Interference than those targeting other regions such as coat protein (CP) sequence. Cas13a can also process pre-crRNAs into functional crRNAs. Our data indicate that CRISPR/Cas13a can be used for engineering Interference against RNA Viruses, providing a potential novel mechanism for RNA-guided immunity against RNA Viruses and for other RNA manipulations in plants.
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rna Virus Interference via crispr cas13a system in plants
bioRxiv, 2017Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Muhammad Zuhaib Khan, Shouwei Ding, Magdy M MahfouzAbstract:CRISPR/Cas systems confer immunity against invading nucleic acids and phages in bacteria and archaea. CRISPR/Cas13a (known previously as C2c2) is a class 2 type VI-A ribonuclease capable of targeting and cleaving single stranded RNA (ssRNA) molecules of the phage genome. Here, we employ CRISPR/Cas13a to engineer Interference with an RNA Virus, Turnip Mosaic Virus (TuMV), in plants. CRISPR/Cas13a produced Interference against green fluorescent protein (GFP) expressing TuMV in transient assays and stable overexpression lines of Nicotiana benthamiana. crRNAs targeting the HC-Pro and GFP sequences exhibited better Interference than those targeting other regions such as coat protein (CP) sequence. Cas13a can also process pre-crRNAs into functional crRNAs. Our data indicate that CRISPR/Cas13a can be used for engineering Interference against RNA Viruses, providing a potential novel mechanism for RNA-guided immunity against RNA Viruses, and for other RNA manipulations in plants.
Rashid Aman - One of the best experts on this subject based on the ideXlab platform.
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crispr cas13d mediates robust rna Virus Interference in plants
Genome Biology, 2019Co-Authors: Ahmed Mahas, Rashid Aman, Magdy M MahfouzAbstract:CRISPR-Cas systems endow bacterial and archaeal species with adaptive immunity mechanisms to fend off invading phages and foreign genetic elements. CRISPR-Cas9 has been harnessed to confer Virus Interference against DNA Viruses in eukaryotes, including plants. In addition, CRISPR-Cas13 systems have been used to target RNA Viruses and the transcriptome in mammalian and plant cells. Recently, CRISPR-Cas13a has been shown to confer modest Interference against RNA Viruses. Here, we characterized a set of different Cas13 variants to identify those with the most efficient, robust, and specific Interference activities against RNA Viruses in planta using Nicotiana benthamiana. Our data show that LwaCas13a, PspCas13b, and CasRx variants mediate high Interference activities against RNA Viruses in transient assays. Moreover, CasRx mediated robust Interference in both transient and stable overexpression assays when compared to the other variants tested. CasRx targets either one Virus alone or two RNA Viruses simultaneously, with robust Interference efficiencies. In addition, CasRx exhibits strong specificity against the target Virus and does not exhibit collateral activity in planta. Our data establish CasRx as the most robust Cas13 variant for RNA Virus Interference applications in planta and demonstrate its suitability for studying key questions relating to Virus biology.
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engineering rna Virus Interference via the crispr cas13 machinery in arabidopsis
Viruses, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Magdy M MahfouzAbstract:Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) systems are key immune mechanisms helping prokaryotic species fend off RNA and DNA Viruses. CRISPR/Cas9 has broad applications in basic research and biotechnology and has been widely used across eukaryotic species for genome engineering and functional analysis of genes. The recently developed CRISPR/Cas13 systems target RNA rather than DNA and thus offer new potential for transcriptome engineering and combatting RNA Viruses. Here, we used CRISPR/LshCas13a to stably engineer Arabidopsis thaliana for Interference against the RNA genome of Turnip mosaic Virus (TuMV). Our data demonstrate that CRISPR RNAs (crRNAs) guiding Cas13a to the sequences encoding helper component proteinase silencing suppressor (HC-Pro) or GFP target 2 (GFP-T2) provide better Interference compared to crRNAs targeting other regions of the TuMV RNA genome. This work demonstrates the exciting potential of CRISPR/Cas13 to be used as an antiviral strategy to obstruct RNA Viruses, and encourages the search for more robust and effective Cas13 variants or CRISPR systems that can target RNA.
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rna Virus Interference via crispr cas13a system in plants
Genome Biology, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Muhammad Zuhaib Khan, Shouwei Ding, Magdy M MahfouzAbstract:CRISPR/Cas systems confer immunity against invading nucleic acids and phages in bacteria and archaea. CRISPR/Cas13a (known previously as C2c2) is a class 2 type VI-A ribonuclease capable of targeting and cleaving single-stranded RNA (ssRNA) molecules of the phage genome. Here, we employ CRISPR/Cas13a to engineer Interference with an RNA Virus, Turnip Mosaic Virus (TuMV), in plants. CRISPR/Cas13a produces Interference against green fluorescent protein (GFP)-expressing TuMV in transient assays and stable overexpression lines of Nicotiana benthamiana. CRISPR RNA (crRNAs) targeting the HC-Pro and GFP sequences exhibit better Interference than those targeting other regions such as coat protein (CP) sequence. Cas13a can also process pre-crRNAs into functional crRNAs. Our data indicate that CRISPR/Cas13a can be used for engineering Interference against RNA Viruses, providing a potential novel mechanism for RNA-guided immunity against RNA Viruses and for other RNA manipulations in plants.
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RNA Virus Interference via CRISPR/Cas13a system in plants
Genome Biology, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Muhammad Zuhaib Khan, Shouwei Ding, Zahir Ali, Magdy MahfouzAbstract:Background CRISPR/Cas systems confer immunity against invading nucleic acids and phages in bacteria and archaea. CRISPR/Cas13a (known previously as C2c2) is a class 2 type VI-A ribonuclease capable of targeting and cleaving single-stranded RNA (ssRNA) molecules of the phage genome. Here, we employ CRISPR/Cas13a to engineer Interference with an RNA Virus, Turnip Mosaic Virus (TuMV), in plants. Results CRISPR/Cas13a produces Interference against green fluorescent protein (GFP)-expressing TuMV in transient assays and stable overexpression lines of Nicotiana benthamiana. CRISPR RNA (crRNAs) targeting the HC-Pro and GFP sequences exhibit better Interference than those targeting other regions such as coat protein (CP) sequence. Cas13a can also process pre-crRNAs into functional crRNAs. Conclusions Our data indicate that CRISPR/Cas13a can be used for engineering Interference against RNA Viruses, providing a potential novel mechanism for RNA-guided immunity against RNA Viruses and for other RNA manipulations in plants.
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Engineering RNA Virus Interference via the CRISPR/Cas13 Machinery in Arabidopsis
MDPI AG, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Zahir Ali, Fatimah Aljedaani, Magdy MahfouzAbstract:Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) systems are key immune mechanisms helping prokaryotic species fend off RNA and DNA Viruses. CRISPR/Cas9 has broad applications in basic research and biotechnology and has been widely used across eukaryotic species for genome engineering and functional analysis of genes. The recently developed CRISPR/Cas13 systems target RNA rather than DNA and thus offer new potential for transcriptome engineering and combatting RNA Viruses. Here, we used CRISPR/LshCas13a to stably engineer Arabidopsis thaliana for Interference against the RNA genome of Turnip mosaic Virus (TuMV). Our data demonstrate that CRISPR RNAs (crRNAs) guiding Cas13a to the sequences encoding helper component proteinase silencing suppressor (HC-Pro) or GFP target 2 (GFP-T2) provide better Interference compared to crRNAs targeting other regions of the TuMV RNA genome. This work demonstrates the exciting potential of CRISPR/Cas13 to be used as an antiviral strategy to obstruct RNA Viruses, and encourages the search for more robust and effective Cas13 variants or CRISPR systems that can target RNA
Ahmed Mahas - One of the best experts on this subject based on the ideXlab platform.
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crispr cas13d mediates robust rna Virus Interference in plants
Genome Biology, 2019Co-Authors: Ahmed Mahas, Rashid Aman, Magdy M MahfouzAbstract:CRISPR-Cas systems endow bacterial and archaeal species with adaptive immunity mechanisms to fend off invading phages and foreign genetic elements. CRISPR-Cas9 has been harnessed to confer Virus Interference against DNA Viruses in eukaryotes, including plants. In addition, CRISPR-Cas13 systems have been used to target RNA Viruses and the transcriptome in mammalian and plant cells. Recently, CRISPR-Cas13a has been shown to confer modest Interference against RNA Viruses. Here, we characterized a set of different Cas13 variants to identify those with the most efficient, robust, and specific Interference activities against RNA Viruses in planta using Nicotiana benthamiana. Our data show that LwaCas13a, PspCas13b, and CasRx variants mediate high Interference activities against RNA Viruses in transient assays. Moreover, CasRx mediated robust Interference in both transient and stable overexpression assays when compared to the other variants tested. CasRx targets either one Virus alone or two RNA Viruses simultaneously, with robust Interference efficiencies. In addition, CasRx exhibits strong specificity against the target Virus and does not exhibit collateral activity in planta. Our data establish CasRx as the most robust Cas13 variant for RNA Virus Interference applications in planta and demonstrate its suitability for studying key questions relating to Virus biology.
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engineering rna Virus Interference via the crispr cas13 machinery in arabidopsis
Viruses, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Magdy M MahfouzAbstract:Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) systems are key immune mechanisms helping prokaryotic species fend off RNA and DNA Viruses. CRISPR/Cas9 has broad applications in basic research and biotechnology and has been widely used across eukaryotic species for genome engineering and functional analysis of genes. The recently developed CRISPR/Cas13 systems target RNA rather than DNA and thus offer new potential for transcriptome engineering and combatting RNA Viruses. Here, we used CRISPR/LshCas13a to stably engineer Arabidopsis thaliana for Interference against the RNA genome of Turnip mosaic Virus (TuMV). Our data demonstrate that CRISPR RNAs (crRNAs) guiding Cas13a to the sequences encoding helper component proteinase silencing suppressor (HC-Pro) or GFP target 2 (GFP-T2) provide better Interference compared to crRNAs targeting other regions of the TuMV RNA genome. This work demonstrates the exciting potential of CRISPR/Cas13 to be used as an antiviral strategy to obstruct RNA Viruses, and encourages the search for more robust and effective Cas13 variants or CRISPR systems that can target RNA.
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rna Virus Interference via crispr cas13a system in plants
Genome Biology, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Muhammad Zuhaib Khan, Shouwei Ding, Magdy M MahfouzAbstract:CRISPR/Cas systems confer immunity against invading nucleic acids and phages in bacteria and archaea. CRISPR/Cas13a (known previously as C2c2) is a class 2 type VI-A ribonuclease capable of targeting and cleaving single-stranded RNA (ssRNA) molecules of the phage genome. Here, we employ CRISPR/Cas13a to engineer Interference with an RNA Virus, Turnip Mosaic Virus (TuMV), in plants. CRISPR/Cas13a produces Interference against green fluorescent protein (GFP)-expressing TuMV in transient assays and stable overexpression lines of Nicotiana benthamiana. CRISPR RNA (crRNAs) targeting the HC-Pro and GFP sequences exhibit better Interference than those targeting other regions such as coat protein (CP) sequence. Cas13a can also process pre-crRNAs into functional crRNAs. Our data indicate that CRISPR/Cas13a can be used for engineering Interference against RNA Viruses, providing a potential novel mechanism for RNA-guided immunity against RNA Viruses and for other RNA manipulations in plants.
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RNA Virus Interference via CRISPR/Cas13a system in plants
Genome Biology, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Muhammad Zuhaib Khan, Shouwei Ding, Zahir Ali, Magdy MahfouzAbstract:Background CRISPR/Cas systems confer immunity against invading nucleic acids and phages in bacteria and archaea. CRISPR/Cas13a (known previously as C2c2) is a class 2 type VI-A ribonuclease capable of targeting and cleaving single-stranded RNA (ssRNA) molecules of the phage genome. Here, we employ CRISPR/Cas13a to engineer Interference with an RNA Virus, Turnip Mosaic Virus (TuMV), in plants. Results CRISPR/Cas13a produces Interference against green fluorescent protein (GFP)-expressing TuMV in transient assays and stable overexpression lines of Nicotiana benthamiana. CRISPR RNA (crRNAs) targeting the HC-Pro and GFP sequences exhibit better Interference than those targeting other regions such as coat protein (CP) sequence. Cas13a can also process pre-crRNAs into functional crRNAs. Conclusions Our data indicate that CRISPR/Cas13a can be used for engineering Interference against RNA Viruses, providing a potential novel mechanism for RNA-guided immunity against RNA Viruses and for other RNA manipulations in plants.
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Engineering RNA Virus Interference via the CRISPR/Cas13 Machinery in Arabidopsis
MDPI AG, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Zahir Ali, Fatimah Aljedaani, Magdy MahfouzAbstract:Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) systems are key immune mechanisms helping prokaryotic species fend off RNA and DNA Viruses. CRISPR/Cas9 has broad applications in basic research and biotechnology and has been widely used across eukaryotic species for genome engineering and functional analysis of genes. The recently developed CRISPR/Cas13 systems target RNA rather than DNA and thus offer new potential for transcriptome engineering and combatting RNA Viruses. Here, we used CRISPR/LshCas13a to stably engineer Arabidopsis thaliana for Interference against the RNA genome of Turnip mosaic Virus (TuMV). Our data demonstrate that CRISPR RNAs (crRNAs) guiding Cas13a to the sequences encoding helper component proteinase silencing suppressor (HC-Pro) or GFP target 2 (GFP-T2) provide better Interference compared to crRNAs targeting other regions of the TuMV RNA genome. This work demonstrates the exciting potential of CRISPR/Cas13 to be used as an antiviral strategy to obstruct RNA Viruses, and encourages the search for more robust and effective Cas13 variants or CRISPR systems that can target RNA
Fatimah R Aljedaani - One of the best experts on this subject based on the ideXlab platform.
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engineering rna Virus Interference via the crispr cas13 machinery in arabidopsis
Viruses, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Magdy M MahfouzAbstract:Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) systems are key immune mechanisms helping prokaryotic species fend off RNA and DNA Viruses. CRISPR/Cas9 has broad applications in basic research and biotechnology and has been widely used across eukaryotic species for genome engineering and functional analysis of genes. The recently developed CRISPR/Cas13 systems target RNA rather than DNA and thus offer new potential for transcriptome engineering and combatting RNA Viruses. Here, we used CRISPR/LshCas13a to stably engineer Arabidopsis thaliana for Interference against the RNA genome of Turnip mosaic Virus (TuMV). Our data demonstrate that CRISPR RNAs (crRNAs) guiding Cas13a to the sequences encoding helper component proteinase silencing suppressor (HC-Pro) or GFP target 2 (GFP-T2) provide better Interference compared to crRNAs targeting other regions of the TuMV RNA genome. This work demonstrates the exciting potential of CRISPR/Cas13 to be used as an antiviral strategy to obstruct RNA Viruses, and encourages the search for more robust and effective Cas13 variants or CRISPR systems that can target RNA.
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Engineering resistance against Tomato yellow leaf curl Virus via the CRISPR/Cas9 system in tomato
Plant signaling & behavior, 2018Co-Authors: Manal Tashkandi, Zahir Ali, Fatimah R Aljedaani, Ashwag Shami, Magdy M MahfouzAbstract:CRISPR/Cas systems confer molecular immunity against phages and conjugative plasmids in prokaryotes. Recently, CRISPR/Cas9 systems have been used to confer Interference against eukaryotic Viruses. Here, we engineered Nicotiana benthamiana and tomato (Solanum lycopersicum) plants with the CRISPR/Cas9 system to confer immunity against the Tomato yellow leaf curl Virus (TYLCV). Targeting the TYLCV genome with Cas9-single guide RNA at the sequences encoding the coat protein (CP) or replicase (Rep) resulted in efficient Virus Interference, as evidenced by low accumulation of the TYLCV DNA genome in the transgenic plants. The CRISPR/Cas9-based immunity remained active across multiple generations in the N. benthamiana and tomato plants. Together, our results confirmed the efficiency of the CRISPR/Cas9 system for stable engineering of TYLCV resistance in N. benthamiana and tomato, and opens the possibilities of engineering Virus resistance against single and multiple infectious Viruses in other crops.
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rna Virus Interference via crispr cas13a system in plants
Genome Biology, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Muhammad Zuhaib Khan, Shouwei Ding, Magdy M MahfouzAbstract:CRISPR/Cas systems confer immunity against invading nucleic acids and phages in bacteria and archaea. CRISPR/Cas13a (known previously as C2c2) is a class 2 type VI-A ribonuclease capable of targeting and cleaving single-stranded RNA (ssRNA) molecules of the phage genome. Here, we employ CRISPR/Cas13a to engineer Interference with an RNA Virus, Turnip Mosaic Virus (TuMV), in plants. CRISPR/Cas13a produces Interference against green fluorescent protein (GFP)-expressing TuMV in transient assays and stable overexpression lines of Nicotiana benthamiana. CRISPR RNA (crRNAs) targeting the HC-Pro and GFP sequences exhibit better Interference than those targeting other regions such as coat protein (CP) sequence. Cas13a can also process pre-crRNAs into functional crRNAs. Our data indicate that CRISPR/Cas13a can be used for engineering Interference against RNA Viruses, providing a potential novel mechanism for RNA-guided immunity against RNA Viruses and for other RNA manipulations in plants.
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RNA Virus Interference via CRISPR/Cas13a system in plants
Genome Biology, 2018Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Muhammad Zuhaib Khan, Shouwei Ding, Zahir Ali, Magdy MahfouzAbstract:Background CRISPR/Cas systems confer immunity against invading nucleic acids and phages in bacteria and archaea. CRISPR/Cas13a (known previously as C2c2) is a class 2 type VI-A ribonuclease capable of targeting and cleaving single-stranded RNA (ssRNA) molecules of the phage genome. Here, we employ CRISPR/Cas13a to engineer Interference with an RNA Virus, Turnip Mosaic Virus (TuMV), in plants. Results CRISPR/Cas13a produces Interference against green fluorescent protein (GFP)-expressing TuMV in transient assays and stable overexpression lines of Nicotiana benthamiana. CRISPR RNA (crRNAs) targeting the HC-Pro and GFP sequences exhibit better Interference than those targeting other regions such as coat protein (CP) sequence. Cas13a can also process pre-crRNAs into functional crRNAs. Conclusions Our data indicate that CRISPR/Cas13a can be used for engineering Interference against RNA Viruses, providing a potential novel mechanism for RNA-guided immunity against RNA Viruses and for other RNA manipulations in plants.
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rna Virus Interference via crispr cas13a system in plants
bioRxiv, 2017Co-Authors: Rashid Aman, Ahmed Mahas, Haroon Butt, Fatimah R Aljedaani, Muhammad Zuhaib Khan, Shouwei Ding, Magdy M MahfouzAbstract:CRISPR/Cas systems confer immunity against invading nucleic acids and phages in bacteria and archaea. CRISPR/Cas13a (known previously as C2c2) is a class 2 type VI-A ribonuclease capable of targeting and cleaving single stranded RNA (ssRNA) molecules of the phage genome. Here, we employ CRISPR/Cas13a to engineer Interference with an RNA Virus, Turnip Mosaic Virus (TuMV), in plants. CRISPR/Cas13a produced Interference against green fluorescent protein (GFP) expressing TuMV in transient assays and stable overexpression lines of Nicotiana benthamiana. crRNAs targeting the HC-Pro and GFP sequences exhibited better Interference than those targeting other regions such as coat protein (CP) sequence. Cas13a can also process pre-crRNAs into functional crRNAs. Our data indicate that CRISPR/Cas13a can be used for engineering Interference against RNA Viruses, providing a potential novel mechanism for RNA-guided immunity against RNA Viruses, and for other RNA manipulations in plants.
Michele Martins - One of the best experts on this subject based on the ideXlab platform.
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Comprehensive Quantitative Proteome Analysis of Aedes aegypti Identifies Proteins and Pathways Involved in Wolbachia pipientis and Zika Virus Interference Phenomenon
Frontiers in physiology, 2021Co-Authors: Michele Martins, Luis Felipe Costa Ramos, Jimmy Rodriguez Murillo, André Q. Torres, Stephanie Serafim De Carvalho, Gilberto B. Domont, Danielle M.p. Oliveira, Rafael D. Mesquita, Fábio C. S. Nogueira, Rafael Maciel-de-freitasAbstract:Zika Virus (ZIKV) is a global public health emergency due to its association with microcephaly, Guillain-Barré syndrome, neuropathy, and myelitis in children and adults. A total of 87 countries have had evidence of autochthonous mosquito-borne transmission of ZIKV, distributed across four continents, and no antiVirus therapy or vaccines are available. Therefore, several strategies have been developed to target the main mosquito vector, Aedes aegypti, to reduce the burden of different arboViruses. Among such strategies, the use of the maternally-inherited endosymbiont Wolbachia pipientis has been applied successfully to reduce Virus susceptibility and decrease transmission. However, the mechanisms by which Wolbachia orchestrate resistance to ZIKV infection remain to be elucidated. In this study, we apply isobaric labeling quantitative mass spectrometry (MS)-based proteomics to quantify proteins and identify pathways altered during ZIKV infection; Wolbachia infection; co-infection with Wolbachia/ZIKV in the A. aegypti heads and salivary glands. We show that Wolbachia regulates proteins involved in reactive oxygen species production, regulates humoral immune response, and antioxidant production. The reduction of ZIKV polyprotein in the presence of Wolbachia in mosquitoes was determined by MS and corroborates the idea that Wolbachia helps to block ZIKV infections in A. aegypti. The present study offers a rich resource of data that may help to elucidate mechanisms by which Wolbachia orchestrate resistance to ZIKV infection in A. aegypti, and represents a step further on the development of new targeted methods to detect and quantify ZIKV and Wolbachia directly in complex tissues.
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comprehensive quantitative proteome analysis of aedes aegypti identifies proteins and pathways involved in wolbachia pipientis and zika Virus Interference phenomenon
Frontiers in Physiology, 2021Co-Authors: Michele Martins, Luis Felipe Costa Ramos, Jimmy Rodriguez Murillo, Stephanie Serafim De Carvalho, Gilberto B. Domont, Danielle M.p. Oliveira, Rafael D. Mesquita, Fábio C. S. Nogueira, Andre Torres, Rafael MacieldefreitasAbstract:Zika Virus (ZIKV) is a global public health emergency due to its association with microcephaly, Guillain-Barre syndrome, neuropathy, and myelitis in children and adults. A total of 87 countries have had evidence of autochthonous mosquito-borne transmission of ZIKV, distributed across four continents, and no antiVirus therapy or vaccines are available. Therefore, several strategies have been developed to target the main mosquito vector, Aedes aegypti, to reduce the burden of different arboViruses. Among such strategies, the use of the maternally-inherited endosymbiont Wolbachia pipientis has been applied successfully to reduce Virus susceptibility and decrease transmission. However, the mechanisms by which Wolbachia orchestrate resistance to ZIKV infection remain to be elucidated. In this study, we apply isobaric labeling quantitative mass spectrometry (MS)-based proteomics to quantify proteins and identify pathways altered during ZIKV infection; Wolbachia infection; co-infection with Wolbachia/ZIKV in the A. aegypti heads and salivary glands. We show that Wolbachia regulates proteins involved in reactive oxygen species production, regulates humoral immune response, and antioxidant production. The reduction of ZIKV polyprotein in the presence of Wolbachia in mosquitoes was determined by MS and corroborates the idea that Wolbachia helps to block ZIKV infections in A. aegypti. The present study offers a rich resource of data that may help to elucidate mechanisms by which Wolbachia orchestrate resistance to ZIKV infection in A. aegypti, and represents a step further on the development of new targeted methods to detect and quantify ZIKV and Wolbachia directly in complex tissues.
