pRGEB32
(Plasmid #63142)

• Purpose: (Empty Backbone) Express sgRNA/PTG with rice snoRNA U3 promoter and Cas9 with rice ubiquitin promoter for Agrobacterium-mediated transformation.
• Depositing Lab: Yinong Yang
• Publication: Xie et al Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2.

Backbone

• Vector backbone: pRGEB31
• Backbone manufacturer: Kabin Xie
• Modifications to backbone: Replace the 35S promoter with rice UBI promoter plus 5'-UTR.
• Vector type: Plant Expression, CRISPR
• Promoter: Rice snoRNA U3 promoter for gRNA/PTG expression and UBI promoter for Cas9 expression
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: M13R(-48)
• 3′ sequencing primer: M13F(-20)

Resource Information

• Supplemental Documents:
  • Annotated Genbank file
• Addgene Notes:
  • Addgene diagnostic digest
  • Addgene Diagnostic Digest
• A portion of this plasmid was derived from a plasmid made by: The Cas9 gene originates from plasmid pX330 created by Dr. Feng Zhang's Lab.
• References:
  • Published Protocol - Targeted Gene Mutation in Rice Using a CRISPR-Cas9 System
  • CRISPR-Plant gRNA spacer design tool
• Articles Citing this Plasmid:
  • 30 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Please use the reference links above to access a published protocol for using plasmids pRGE31, pRGEB31, pRGE32 and pRGEB32 as well as a CRISPR design software tool.