Conferring Plant Resistance to Geminiviruses using CRISPR/Cas Systems
CRISPR/Cas Systems Provide Plants with Geminivirus ResistanceCRISPR/Cas systems can be used to create geminivirus resistance in plants. Using this genome editing tool, plants are pre-programmed with a Cas endonuclease and small RNA sequence that essentially give them an immune system that targets both conserved and non-conserved nucleotide sequences within the geminivirus genome. The technology can inhibit the geminivirus life cycle in three ways: 1) a Cas endonuclease creates targeted DNA double-strand breaks in the geminivirus genome, 2) nickase versions of Cas create targeted DNA nicks in both strands of the geminivirus genome, 3) a nuclease-dead version of Cas blocks the virus or host proteins from functioning. Additionally, for each approach, multiple regions on one or more Geminiviruses can be targeted simultaneously. By actually destroying the geminivirus DNA, this technique is expected to be highly effective at preventing geminivirus disease.Destroys the Geminivirus GenomeCurrent approaches to generating geminivirus resistant plants include expressing geminivirus coat proteins, replication genes or antisense RNAs, as well as using defective-interfering replicons. However, such efforts have had limited success. This CRISPR/Cas technology improves upon current methods in several aspects; most importantly, it destroys the geminivirus genome in order to prevent disease. The technology can be multiplexed to target multiple regions of either the same virus or multiple viruses, thus enabling longer-lasting resistance. Furthermore, it can be optimized differently (i.e. as an endonuclease, nickase or a physical blockade) in different plant species.BENEFITS AND FEATURES:CRISPR/Cas systems destroy foreign viral DNA, thereby preventing diseaseCan be multiplexed to target multiple regions of the virus and enable longer-lasting resistanceVersatile, and can be optimized differently in different plant speciesNuclease-dead versions of endonucleases physically block the virus or host proteins from functioningEase of reprogramming permits most researchers with basic molecular biology experience to apply the technologyAPPLICATIONS:Maize Streak MastrevirusAfrican Cassava Mosaic VirusAny crop or economically valuable plant susceptible to geminiviruses that is amenable to stable DNA integration, especially African cropsPhase of Development - Proof of Concept
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