pRGE31
(Plasmid #50929)

• Purpose: (Empty Backbone) Expressing sgRNA and Cas9 in plants. The sgRNA was controlled by rice snoRNA U3 promoter
• Depositing Lab: Yinong Yang
• Publication: Xie et al Mol Plant. 2013 Nov;6(6):1975-83. doi: 10.1093/mp/sst119. Epub 2013 Aug 17.

Backbone

• Vector backbone: pUGW11
• Backbone manufacturer: Nakagawa,T.
• Backbone size (bp): 6034
• Modifications to backbone: 1. Removed Bsa I sites. 2. Insert a fragment containing Rice U3 promoter and gRNAscaffold 3. Insert Cas9 by LR reaction.
• Vector type: Plant Expression, CRISPR
• Promoter: Rice snoRNA U3 and dual 35S promoter

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 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

• A portion of this plasmid was derived from a plasmid made by: The Cas9 fragment was from pX330.
• References:
  • Published Protocol - Targeted Gene Mutation in Rice Using a CRISPR-Cas9 System
  • CRISPR-Plant gRNA spacer design tool

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.