Efficient and modular CRISPR-Cas9 vector system for Physcomitrella patens.

Mallett DR, Chang M, Cheng X, Bezanilla M
Plant Direct. 2019 Sep 12;3(9):e00168. doi: 10.1002/pld3.168. eCollection 2019 Sep. PubMed Article

These materials are included in the CRISPR-Cas9 Gateway System for Physcomitrella patens Kit.

Plasmids from Article

ID	Plasmid	Purpose
113735	pENTR-PpU6P-sgRNA-L1L2	Gateway entry vector containing attL1 and attL2 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
113736	pENTR-PpU6P-sgRNA-L1L4	Gateway entry vector containing attL1 and attL4 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
113737	pENTR-PpU6P-sgRNA-L1R5	Gateway entry vector containing attL1 and attR5 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
113738	pENTR-PpU6P-sgRNA-L5L2	Gateway entry vector containing attL5 and attL2 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
113739	pENTR-PpU6P-sgRNA-R4R3	Gateway entry vector containing attR4 and attR3 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
113740	pENTR-PpU6P-sgRNA-L3L2	Gateway entry vector containing attL3 and attL2 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
113741	pENTR-PpU6P-sgRNA-L5L4	Gateway entry vector containing attL5 and attL4 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
113742	pMH-Cas9-gate	Gateway destination vector encoding Maize ubiquitin promoter driving Cas9 and 35S promoter driving hygromycin for selection in plants.
113743	pMK-Cas9-gate	Gateway destination vector encoding Maize ubiquitin promoter driving Cas9 and 35S promoter driving aminoglycoside phosphotransferase for G418 selection in plants.
113744	pZeo-Cas9-gate	Gateway destination vector encoding Maize ubiquitin promoter driving Cas9 and 35S promoter driving Zeocin for selection in plants.
113745	pENTR-R4R3-stop	Contains three stop codons in every frame to ensure a premature stop codon when introduced into the genome via homology-directed repair. Gateway multisite entry clone.
113746	pENTR-R4R3-mEGFP-N	Gateway multi-site entry clone for second position mEGFP (N-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
113747	pENTR-R4R3-2xmEGFP-N	Gateway multi-site entry clone for second position 2x mEGFP (N-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
113748	pENTR-R4R3-3xmEGFP-N	Gateway multi-site entry clone for second position 3x mEGFP (N-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
113749	pENTR-R4R3-mEGFP-C	Gateway multi-site entry clone for second position mEGFP (C-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
113750	pENTR-R4R3-2xmEGFP-C	Gateway multi-site entry clone for second position 2x mEGFP (C-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
113751	pENTR-R4R3-3xmEGFP-C	Gateway multi-site entry clone for second position 3x mEGFP (C-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
113752	pENTR-R4R3-mRuby-N	Gateway multi-site entry clone for second position mRuby (N-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
113753	pENTR-R4R3-2xmRuby-N	Gateway multi-site entry clone for second position 2x mRuby (N-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
113754	pENTR-R4R3-3xmRuby-N	Gateway multi-site entry clone for second position 3x mRuby (N-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
113755	pENTR-R4R3-mRuby-C	Gateway multi-site entry clone for second position mRuby (C-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
113756	pENTR-R4R3-2xmRuby-C	Gateway multi-site entry clone for second position 2x mRuby (C-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
113757	pENTR-R4R3-3xmRuby-C	Gateway multi-site entry clone for second position 3x mRuby (C-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
113758	pGEM-gate	Gateway destination vector only containing the Gateway cassette and pGEM-T Easy backbone. This can be used with Gateway-compatible Homology-Directed Repair.