gRNA NIA TLS1
(Plasmid #196980)

• Purpose: Expresses two gRNAs targeting Arabidopsis NIA1 gene for knockout. Each gRNA is fused to TLS1 mobility signal that confers graft mobility.
• Depositing Lab: Friedrich Kragler
• Publication: Yang et al Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01585-8.

Backbone

• Vector backbone: pMDC100
• Total vector size (bp): 11136
• Vector type: Plant Expression
• Selectable markers: Kanamycin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: AtNIA1 gRNAs fused to TLS1 tags
• gRNA/shRNA sequence: ACAACACTGCTGACTCTGCA, GATGGGTTACAACGTGAAGG
• Species: A. thaliana (mustard weed); Streptococcus pyogenes
• Promoter: pU6-26, pU6-29

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsaI (destroyed during cloning)
• 3′ cloning site: BsaI (destroyed during cloning)
• 5′ sequencing primer: CCAATTCAGTCGACTGGATCC
• 3′ sequencing primer: ggcggccgctctagaactag

Resource Information

• Supplemental Documents:
  • grna-nia1-tls1.gb
  • GO CRISP Cloning Protocol

Terms and Licenses

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

Depositor Comments

Please refer to the GO CRISP Cloning Protocol linked above for additional information.