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.
Original language | English (US) |
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Article number | 1 |
Journal | Genome biology |
Volume | 19 |
Issue number | 1 |
DOIs | |
State | Published - 2018 |
Keywords
- CRISPR/Cas systems
- CRISPR/Cas13a
- Molecular immunity
- RNA interference
- RNA knockdown
- Transcriptome regulation
- Virus interference
ASJC Scopus subject areas
- Ecology, Evolution, Behavior and Systematics
- Genetics
- Cell Biology
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RNA virus interference via CRISPR/Cas13a system in plants
Aman, R. (Creator), Ali, Z. (Creator), Butt, H. (Creator), Mahas, A. (Creator), Aljedaani, F. R. (Creator), Khan, M. Z. (Creator), Ding, S. (Creator), Mahfouz, M. M. (Creator), Khan, M. Z. (Creator) & Ding, S. (Creator), figshare, 2018
DOI: 10.6084/m9.figshare.c.3968844.v1, http://hdl.handle.net/10754/664094
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