Genome editing vectors
Last modified October 2, 2014
Reference:
•Jacobs, TB, PR LaFayette, MA Ortega, RJ Schmitz, WA Parrott. 2015. Targeted genome modifications in soybean with CRISPR/Cas9. BMC Biotechnology. 15:16. PDF
What is our CRISPR vector family?
•The CRISPR family is a series of binary vectors for plant transformation. The selectable markers include GFP, bar, hygromycin, and nptII. Additional guide RNAs can be introduced with the meganuclease I-PpoI.
•In our hairy-root system, >95% of events contain modifications and >70% of alleles are modified.
Knockout of GFP with CRISPR in soybean:
Protocols for CRISPR vectors:
Where can I obtain the CRISPR family vectors?
Click the plasmid names maps to visit their addgene page for downloading their .gb files
| Addgene ID | Plasmid | Purpose |
|---|---|---|
| 59178 | p201G Cas9 | Cas9 driven by double 35S, GFP for plant selection, I-PpoI site to accept gRNA from pUC gRNA Shuttle |
| 59177 | p201B Cas9 | Cas9 driven by double 35S, BAR for plant selection, I-PpoI site to accept gRNA from pUC gRNA Shuttle |
| 59176 | p201H Cas9 | Cas9 driven by double 35S, hygromycin resistance for plant selection, I-PpoI site to accept gRNA from pUC gRNA Shuttle |
| 59175 | p201N Cas9 | Cas9 driven by double 35S, nptII for plant selection, I-PpoI site to accept gRNA from pUC gRNA Shuttle |
| 47024 | pUC gRNA Shuttle | Encodes a template from which gRNAs can be made via InFusion cloning. The Medicago truncatula U6.6 promoter drives the gRNA. For use in plants. |