A modular toolbox for gRNA-Cas9 genome engineering in plants based on the GoldenBraid standard.
Vazquez-Vilar M, Bernabe-Orts JM, Fernandez-Del-Carmen A, Ziarsolo P, Blanca J, Granell A, Orzaez D
Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. eCollection 2016. PubMed
Vazquez-Vilar M, Bernabe-Orts JM, Fernandez-Del-Carmen A, Ziarsolo P, Blanca J, Granell A, Orzaez D
Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. eCollection 2016. PubMed
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 75399 | pdCas9 (GB1079) | Provides the human codon optimized CDS of Cas9 protein with mutated (D10A, H840A) and inactivated catalytic domains as a level 0 GoldenBraid part for C-terminal fusions |
| 75400 | pEGB2Ω2_SF-35s:hCas9:tNos (GB1103) | Transcriptional unit for (human codon optimized) Cas9 plant expression driven by the 35S promoter. Specially conceived to be linked with the TU of the kanamycin resistance gene GB1181. |
| 75401 | pEGB 35s:dCas:BRD:tNos (GB1172) | Transcriptional unit of (human codon optimized) inactivated Cas9 fused to the BRD Transcriptional Repressor |
| 75403 | pEGB 35s:dCas:VP64:tNos (GB1189) | Transcriptional unit of (human codon optimized) inactivated Cas9 fused to the VP64 Transcriptional Activator |
| 75404 | pEGB 35s:dCas:EDLL:tNos (GB1190) | Transcriptional unit of (human codon optimized) inactivated Cas9 fused to the EDLL Transcriptional Activator |
| 75405 | pU6-1 (GB1204) | Provides the A. thaliana U6-1 RNA polIII promoter as a level 0 GoldenBraid part |
| 75406 | tRNA-gRNA position [D1_2] (GB1205) | tRNA and scaffold for the assembly of GBoligomers for the first position (positon [D1_2]) of a polycistronic tRNA-gRNA regulated by the dicot U6-26 or U6-1 promoter (3-part multiplexing) |
| 75407 | tRNA-gRNA position [2_n-1] (GB1206) | tRNA and scaffold for the assembly of GBoligomers for the intermediate position (positon [2_n-1]) of a polycistronic tRNA-gRNA (3-part multiplexing) |
| 75408 | tRNA-gRNA position [n] (GB1207) | tRNA and scaffold for the assembly of GBoligomers for the last position (positon [n]) of a polycistronic tRNA-gRNA (2- and 3-part multiplexing) |
| 75409 | tRNA-gRNA position [D1_n-1] (GB1208) | tRNA and scaffold for the assembly of GBoligomers for the first position (positon [D1_n-1]) of a polycistronic tRNA-gRNA regulated by the U6-26 or U6-1 promoter (2-part multiplexing) |
| 75410 | tRNA-gRNA position [M1_2] (GB1209) | tRNA and scaffold for the assembly of GBoligomers for the first position (positon [M1_2]) of a polycistronic tRNA-gRNA regulated by the (monocot) U3 promoter (3-part multiplexing) |
| 75411 | tRNA-gRNA position [M1_n-1] (GB1210) | tRNA and scaffold for the assembly of GBoligomers for the first position (positon [M1_n-1]) of a polycistronic tRNA-gRNA regulated by the (monocot) U3 promoter (2-part multiplexing) |
| 75412 | pEGB SF-35S:Renilla:TNOS-35S:P19:TNOS (GB0160) | Module for the expression of the Renilla Luciferase with the silencing suppressor P19 |
| 75471 | pEGB3omega1R Tnos:NptII:Pnos-SF (GB1181) | Transcriptional unit for kanamycin resistance gene |