RNA virus interference via CRISPR/Cas13a system in plants

• Published in: Genome Biology Vol. 19; no. 1; p. 1
• Main Authors: Aman, Rashid, Ali, Zahir, Butt, Haroon, Mahas, Ahmed, Aljedaani, Fatimah, Khan, Muhammad Zuhaib, Ding, Shouwei, Mahfouz, Magdy
• Format: Journal Article
• Language: English
• Published: England BioMed Central 04.01.2018; BMC
• Subjects: Amino acid sequence; Coat protein; CRISPR; CRISPR-Associated Proteins - metabolism; CRISPR-Cas Systems; CRISPR/Cas13a; Crop diseases; E coli; Flowers & plants; Genes; Genetic Engineering; Genomes; Green fluorescent protein; Green Fluorescent Proteins - genetics; Molecular immunity; Nicotiana - genetics; Nicotiana - metabolism; Phages; Plant diseases; Plant resistance; Plant viruses; Plasmids; Potyvirus - genetics; Proteins; Ribonucleases - metabolism; RNA - metabolism; RNA Interference; RNA knockdown; RNA viruses; RNA-mediated interference; Target recognition; Transgenic plants; Viral infections; Virus interference; Viruses; Molecular immunity; RNA knockdown; RNA interference; Virus interference; Transcriptome regulation; CRISPR/Cas systems; CRISPR/Cas13a
• ISSN: 1474-760X; 1474-7596; 1474-760X
• DOI: 10.1186/s13059-017-1381-1

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.