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Table 1 Published standard parts for plants for use in the assembly of binary plasmid vectors for Cas9-mediated targeted mutagenesis

From: Meeting report: GARNet/OpenPlant CRISPR-Cas workshop

Name Description AddGene code Publication
Constitutive promoters (PROM) and strong 5′ untranslated regions (5UTR) for use in Cas9 and selectable markers gene cassettes
 pICH51266 PROM (1.3 kb), 35s (Cauliflower Mosaic Virus) + 5UTR omega (Tobacco Mosaic Virus) #50267 [20]
 pICH51288 PROM (double), 35s (Cauliflower Mosaic Virus) + 5UTR, omega (Tobacco Mosaic Virus) #50269 [20]
 pICSL12009 PROM and 5UTR Ubiquitin (Zea mays) #68257 [22]
 pICH87633 PROM, nopoline syntase (Agrobacterium tumefaciens) + 5UTR, omega (Tobacco Mosaic Virus) #50271 [20]
 pICH85281 PROM + 5UTR, mannopine synthase (A. tumefaciens) + 5UTR, omega (Tobacco Mosaic Virus) #50272 [20]
 pICSL12006 PROM + 5UTR (Cassava Vein Mosaic Virus) #50270 [20]
Small RNA promoters for single guide RNAs
 pICSL9003 PROM U6 (Triticum aestivum) #68262 [22]
 pICSL90002 PROM U6-26 (Arabidopsis thaliana) #68261 [22]
Coding sequences (CDS) of selectable markers genes
 pICSL80037 CDS neomycin phosphotransferase II (Escherichia coli) #68260 [22]
 pICSL80036 CDS hygromycin phophotransferase II (E. coli) #68259 [22]
 piCH42222 Phosphinothricin acetyl transferase, (Streptomyces hygroscopicus) #50328 [20]
 pICH43844 CDS phosphinothricin acetyl transferase, (S. hygroscopicus) with intron from ACT2 (A. thaliana) #50329 [20]
3′ untranslated regions (3UTR) and polyadenylation signals (TERM) for use in Cas9 and selectable markers gene cassettes
 pICH41414 3UTR, polyadenylation signal/terminator, 35s (Cauliflower Mosaic Virus) #50337 [20]
 pICH41421 3UTR + TERM, nopaline synthase (A. tumefaciens) #50339 [20]
 pICH41432 3UTR + TERM octopine synthase (A. tumefaciens) #50343 [20]
 piCH72400 3UTR + TERM g7 (A. tumefaciens) #50338 [20]
Cas9 coding sequence and single guide RNA scaffold
 pICH41308 ::hCas9 CDS Cas9 (Streptococcus pyogenes) #49770 [23]
 pICSL11061 Single guide RNA scaffold #46966 [24]
Assembled selectable marker and Cas9 transcriptional units
 pICSL11055 pICH51288 (double 35s) + pICH80037 (nptII) + pICH41421 (nos) in pICH47732 (Level 1 Position 1) backbone #68252 [22]
 pICSL11059 pICH51266 (35s) + PICSL80036 (hptII) + pICH41414 (35 s) in pICH47732 (Level 1 Position 1) backbone #68263 [22]
 pICSL11060 pICSL12006 (CsVMV) + pICH41308::hCas9 (Cas9) + pICH41414 (35s) in pICH47742 (Level 1 Position 2) backbone #68264 [22]
 pICSL11056 pICSL12009 (ZmUbi) + pICH41308::hCas9 (Cas9) + pICH41421(nos) in pICH47742 (Level 1 Position 2) backbone #68258 [22]
 pICH47742::2x35S-5′UTR-hCas9(STOP)-NOST pICH51288 (double 35 s) + pICH41308::hCas9 (Cas9) + pICH41421(nos) in pICH47742 (Level 1 Position 2) backbone #49771 [23]
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Plant Methods

ISSN: 1746-4811