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Genome Biol. 2018 Jan 4;19(1):1. doi: 10.1186/s13059-017-1381-1.

RNA virus interference via CRISPR/Cas13a system in plants.

Author information

1
Laboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and Technology, Thuwal, 23955-6900, Saudi Arabia.
2
Center for Plant Cell Biology, Department of Microbiology and Plant Pathology, University of California, Riverside, CA, 92521, USA.
3
Laboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and Technology, Thuwal, 23955-6900, Saudi Arabia. magdy.mahfouz@kaust.edu.sa.

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.

KEYWORDS:

CRISPR/Cas systems; CRISPR/Cas13a; Molecular immunity; RNA interference; RNA knockdown; Transcriptome regulation; Virus interference

PMID:
29301551
PMCID:
PMC5755456
DOI:
10.1186/s13059-017-1381-1
[Indexed for MEDLINE]
Free PMC Article

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