CRISPR/Cas9-Mediated Viral Interference in Plants

The CRISPR/Cas9 system provides bacterial and archaeal species with molecular immunity against invading phages and conjugative plasmids. The CRISPR/Cas9 has been used for targeted genome editing in diverse eukaryotic species including plants. We investigated whether the CRISPR/Cas9 system could be used in plants to confer molecular immunity against DNA viruses. Indeed, our data show that the CRISPR/Cas9 system is portable to plants. CRISPR/Cas9 machinery including sgRNAs targeting the coding and non-coding sequences of Tomato yellow leaf curl virus (TYLCV) targeted the virus for degradation and introduced mutations at the target sequences. Using a single sgRNA targeting a conserved sequence in Geminiviruses resulted in a simultaneous viral interference against multiple viruses. Taken together, our data establish the efficacy of the CRISPR/Cas9 system for viral interference in plants for addressing basic questions and for biotechnological applications.