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RNA virus interference via CRISPR/Cas13a system in plants.

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.

SUBMITTER: Aman R

PROVIDER: S-EPMC5755456 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature

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RNA virus interference via CRISPR/Cas13a system in plants.

Aman Rashid R Ali Zahir Z Butt Haroon H Mahas Ahmed A Aljedaani Fatimah F Khan Muhammad Zuhaib MZ Ding Shouwei S Mahfouz Magdy M

Genome biology 20180104 1

<h4>Background</h4>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.<h4>Results</h4>CRISPR/Cas13a produces interference against green fluorescent protein (GFP)-expre ...[more]

PMID: 29301551

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