RNA virus interference via CRISPR/Cas13a system in plants
Abstract 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...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2018-01-01
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| Series: | Genome Biology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13059-017-1381-1 |
| _version_ | 1819175003301609472 |
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| author | Rashid Aman Zahir Ali Haroon Butt Ahmed Mahas Fatimah Aljedaani Muhammad Zuhaib Khan Shouwei Ding Magdy Mahfouz |
| author_facet | Rashid Aman Zahir Ali Haroon Butt Ahmed Mahas Fatimah Aljedaani Muhammad Zuhaib Khan Shouwei Ding Magdy Mahfouz |
| author_sort | Rashid Aman |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-22T20:47:57Z |
| format | Article |
| id | doaj.art-14d9462ecf18439d943b2d311e3d545d |
| institution | Directory Open Access Journal |
| issn | 1474-760X |
| language | English |
| last_indexed | 2024-12-22T20:47:57Z |
| publishDate | 2018-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Genome Biology |
| spelling | doaj.art-14d9462ecf18439d943b2d311e3d545d2022-12-21T18:13:10ZengBMCGenome Biology1474-760X2018-01-011911910.1186/s13059-017-1381-1RNA virus interference via CRISPR/Cas13a system in plantsRashid Aman0Zahir Ali1Haroon Butt2Ahmed Mahas3Fatimah Aljedaani4Muhammad Zuhaib Khan5Shouwei Ding6Magdy Mahfouz7Laboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and TechnologyLaboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and TechnologyLaboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and TechnologyLaboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and TechnologyLaboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and TechnologyLaboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and TechnologyCenter for Plant Cell Biology, Department of Microbiology and Plant Pathology, University of CaliforniaLaboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and TechnologyAbstract 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.http://link.springer.com/article/10.1186/s13059-017-1381-1CRISPR/Cas systemsCRISPR/Cas13aVirus interferenceRNA interferenceMolecular immunityRNA knockdown |
| spellingShingle | Rashid Aman Zahir Ali Haroon Butt Ahmed Mahas Fatimah Aljedaani Muhammad Zuhaib Khan Shouwei Ding Magdy Mahfouz RNA virus interference via CRISPR/Cas13a system in plants Genome Biology CRISPR/Cas systems CRISPR/Cas13a Virus interference RNA interference Molecular immunity RNA knockdown |
| title | RNA virus interference via CRISPR/Cas13a system in plants |
| title_full | RNA virus interference via CRISPR/Cas13a system in plants |
| title_fullStr | RNA virus interference via CRISPR/Cas13a system in plants |
| title_full_unstemmed | RNA virus interference via CRISPR/Cas13a system in plants |
| title_short | RNA virus interference via CRISPR/Cas13a system in plants |
| title_sort | rna virus interference via crispr cas13a system in plants |
| topic | CRISPR/Cas systems CRISPR/Cas13a Virus interference RNA interference Molecular immunity RNA knockdown |
| url | http://link.springer.com/article/10.1186/s13059-017-1381-1 |
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