pcU6_3 MS2 sgRNA
(Plasmid #92394)

• Purpose: Chick-specific U6 sgRNA expression mini-vector, harbouring chick U6_3 pol III promoter, with tracrRNA scaffold containing stem loops allowing binding of bacteriophage MS2 coat protein MCP
• Depositing Lab: Tatjana Sauka-Spengler
• Publication: Williams et al Development. 2018 Feb 23;145(4). pii: dev.160333. doi: 10.1242/dev.160333.

Backbone

• Vector backbone: pNG3
• Backbone size w/o insert (bp): 3055
• Total vector size (bp): 3513
• Vector type: Mammalian Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Tetracycline, 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: chick U6.3 promoter and gRNA cloning cassette including MS2 stem loops
• Species: G. gallus (chicken)
• Insert Size (bp): 458
• Promoter: chick U6.3

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: gatcaagcctgattgggagaa

Resource Information

• A portion of this plasmid was derived from a plasmid made by: chick U6.3 promoter (Kudo and Sutou, 2005) was cloned into BsaI sites in pNG3 vector #30987. MS2 stem loops for MCP binding were amplified from #61424

Terms and Licenses

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

Depositor Comments

Kudo, T., and S. Sutou. 2005. Usage of putative chicken U6 promoters for vector-based RNA interference. J Reprod Dev. 51:411-417.

Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic Acids Res. 2011 Apr 14. ():. 10.1093/nar/gkr218 PubMed 21493687

Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F. Nature. 2014 Dec 10. doi: 10.1038/nature14136. 10.1038/nature14136 PubMed 25494202

Please visit https://www.biorxiv.org/content/early/2017/05/08/135525 for bioRxiv preprint.