Generation of virus‐resistant potato plants by RNA genome targeting

Abstract

Summary CRISPR /Cas systems provide bacteria and archaea with molecular immunity against invading phages and foreign plasmids. The class 2 type VI CRISPR /Cas effector Cas13a is an RNA ‐targeting CRISPR effector that provides protection against RNA phages. Here we report the repurposing of CRISPR /Cas13a to protect potato plants from a eukaryotic virus, Potato virus Y ( PVY ). Transgenic potato lines expressing Cas13a/sg RNA (small guide RNA ) constructs showed suppressed PVY accumulation and disease symptoms. The levels of viral resistance correlated with the expression levels of the Cas13a/sg RNA construct in the plants. Our data further demonstrate that appropriately designed sg RNA s can specifically interfere with multiple PVY strains, while having no effect on unrelated viruses such as PVA or Potato virus S . Our findings provide a novel and highly efficient strategy for engineering crops with resistances to viral diseases.

Aphid Transmission of Potyvirus: The Largest Plant-Infecting RNA Virus Genus

“…This study demonstrates the dual benefit of using CRISPR/Cas13a system: conferring resistance to RNA plant viruses by specifically targeting any RNA transcript and, most importantly, blocking the aphid transmission of potyviruses by successful knocking down of HC-Pro. Zhan et al [ 167 ] recently developed transgenic potato lines expressing Cas13a/sgRNA (small guide RNA) constructs, which suppressed PVY accumulation and disease symptoms. Other successful attempts involve engineering transgene-free genetic resistance to potyviruses: TuMV [ 168 ] and clover yellow vein virus (ClYVV) [ 169 ] in Arabidopsis and cassava brown streak virus (CBSV) in cassava [ 170 ], using CRISPR-based technologies specifically targeting eIF4E, a eukaryotic translation initiation factor.…”

CRISPR-Cas13d mediates robust RNA virus interference in plants

“…In addition, we demonstrated that LshCas13a can process pre-crRNA to generate mature and functional crRNAs that guide LshCas13a to degrade the targeted virus. Subsequently, LshCas13a has become the prominent Cas13 orthologue in engineering immunity against different RNA viruses in different plant species, including monocot and dicot plants [34,35]. Since the first report of LshCas13a, several studies have identified more variants of Cas13 proteins belonging to different Cas13 families, which have been classified into four type VI subtypes (A-D) [36][37][38].…”

Control of Plant Viruses by CRISPR/Cas System-Mediated Adaptive Immunity

“…Similarly, resistance to TMV, rice stripe mosaic virus (RSMV), and Southern rice black-streaked dwarf virus (SRBSDV) in transgenic tobacco and rice harboring LshCas13a specifically targeting viral RNA was also confirmed using this system (Zhang et al, 2019a). By targeting the P3-, NIb-, or CP-coding sequences in the potato virus Y (PVY) genomic region, this CRISPR/Cas13a system also showed its effectiveness in interfering with and inhibiting PVY infection (Zhan et al, 2019).…”