title: CRISPR/Cas9-Mediated Immunity to Geminiviruses: Differential Interference and Evasion.
abstract: The CRISPR/Cas9 system has recently been used to confer molecular immunity against several eukaryotic viruses, including plant DNA geminiviruses. Here, we provide a detailed analysis of the efficiencies of targeting different coding and non-coding sequences in the genomes of multiple geminiviruses. Moreover, we analyze the ability of geminiviruses to evade the CRISPR/Cas9 machinery. Our results demonstrate that the CRISPR/Cas9 machinery can efficiently target coding and non-coding sequences and interfere with various geminiviruses. Furthermore, targeting the coding sequences of different geminiviruses resulted in the generation of viral variants capable of replication and systemic movement. By contrast, targeting the noncoding intergenic region sequences of geminiviruses resulted in interference, but with inefficient recovery of mutated viral variants, which thus limited the generation of variants capable of replication and movement. Taken together, our results indicate that targeting noncoding, intergenic sequences provides viral interference activity and significantly limits the generation of viral variants capable of replication and systemic infection, which is essential for developing durable resistance strategies for long-term virus control.
authors.affiliation: Laboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi Arabia.
authors.forename: Zahir
authors.initials: Z
authors.lastname: Ali
authors.affiliation: Laboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi Arabia.
authors.forename: Shakila
authors.initials: S
authors.lastname: Ali
authors.affiliation: Laboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi Arabia.
authors.forename: Manal
authors.initials: M
authors.lastname: Tashkandi
authors.affiliation: Laboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi Arabia.
authors.forename: Syed Shan-E-Ali
authors.initials: SS
authors.lastname: Zaidi
authors.affiliation: Laboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi Arabia.
authors.forename: Magdy M
authors.initials: MM
authors.lastname: Mahfouz
mesh_terms: D001483:Base Sequence; D064113:CRISPR-Cas Systems; D059766:DNA End-Joining Repair; D060467:Disease Resistance; D017791:Geminiviridae / Q000235:genetics* / Q000276:immunology; D005814:Genes, Viral; D054643:INDEL Mutation; D055029:Inverted Repeat Sequences; D016366:Open Reading Frames; D010935:Plant Diseases / Q000276:immunology / Q000821:virology*; D014026:Nicotiana / Q000235:genetics / Q000276:immunology / Q000821:virology; D014764:Viral Proteins / Q000235:genetics
chemical_list: D014764:Viral Proteins