RNA virus interference via CRISPR/Cas13a system in plants

Rashid Aman, Zahir Ali, Haroon Butt, Ahmed Mahas, Fatimah R. Aljedaani, Muhammad Zuhaib Khan, Shouwei Ding, Magdy M. Mahfouz

Research output: Contribution to journalArticlepeer-review

138 Scopus citations

Abstract

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.
Original languageEnglish (US)
JournalGenome biology
Volume19
Issue number1
DOIs
StatePublished - Jan 1 2018

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