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Kamoun Lab CRISPR Plasmids Available from Addgene

A CRISPR/Cas system designed and optimized for work in the model plant Nicotiana benthamiana .

Please refer to the Supplementary methods section of the manuscript for the protocol on assembling customised sgRNA expressing plasmids. Questions regarding the protocol can be addressed to [email protected] or [email protected] .

These plasmids are described in:

Targeted mutagenesis in the model plant Nicotiana benthamiana using Cas9 RNA-guided endonuclease. Nekrasov V, Staskawicz B, Weigel D, Jones JD, Kamoun S. Nat Biotechnol. 2013 Aug 8;31(8):691-3. PubMed PMID 23929340.

Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas system. Belhaj K, Chaparro-Garcia A, Kamoun S, Nekrasov V. Plant Methods. 2013 Oct 11;9(1):39. PubMed PMID 24112467.

Individual plasmids can be ordered via the links below:

ID Plasmid
46965 pK7WGF2::hCas9: Expesses the human codon usage Cas9 nuclease (Mali et al. Science 339, 823-826, 2013) with an N-terminal GFP tag from the 35S promoter in the plant tissue.
46968 pICSL01009::AtU6p: Encodes the Arabidopsis U6 promoter in a level 0 vector (Weber et al. PLoS One 6:e16765, 2011). It is used to place an sgRNA uder the Arabidopsis U6 promoter.
46966 pICH86966::AtU6p::sgRNA_PDS: Expresses an sgRNA targeting the PDS gene in Nicotiana benthamiana from the Arabidopsis U6 promoter.
48075 pICH86966: A binary vector for plant gene expression (S. Marillonnet, described in Weber et al. (PLoS One 6:e16765, 2011).
49770 pICH41308::hCas9: Level 0 hCas9 module.
49771 pICH47742::2x35S-5'UTR-hCas9(STOP)-NOST: Level 1 hCas9 module.
49772 pAGM4723::AtU6p::sgRNA2-2x35S-5'UTR::Cas9::NOST-AtU6p::sgRNA1 : Level 2 construct.