Jun lab tunable CRISPRi strain
(Bacterial strain #86400)

• Purpose: Strain SJ_XTL219 Genotype: MG1655 PBAD_dcas9 ΔlacI ΔaraE araFGH<>spec lacY A177C, galM<pBBa-J23119-tet-sacB-handle-term
• Depositing Lab: Suckjoon Jun
• Publication: Li et al Sci Rep. 2016 Dec 20;6:39076. doi: 10.1038/srep39076.

Backbone

• Vector backbone: strain: SJ_XTL219
• Modifications to backbone: genotype: MG1655 PBAD_dcas9 ΔlacI ΔaraE araFGH<>spec lacY A177C, galM<pBBa-J23119-tet-sacB-handle-terminator>gmpA /pSIM18 hygroR
• Vector type: CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin and hygromycin
• Growth Temperature: 30°C
• Growth Strain(s): MG1655
• Growth instructions: Strain should be grown in 32 oC to maintain pSIM18 plasmid with Hygro.
• Copy number: Unknown

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

THIS IS A STRAIN.
SJ_XTL219 is an E.coli K12 derivative of MG1655. It contains a tunable arabinose operon promoter PBAD to quantitatively control the expression of CRISPR-dCas9 protein over two orders of magnitude in a plasmid-free system. This tCRISPRi is reversible, and gene expression can be repressed or restored by adding or withdrawing arabinose. This tCRISPRi strain has less than 10% leaky expression. Most importantly from a practical perspective, construction of tCRISPRi to target a new gene requires only one-step of oligo recombineering to add the sgRNA. This tCRISPRi strain, in combination with recombineering, provides a simple and easy-to-implement tool for gene expression control, and is ideally suited for construction of both individual strains and high-throughput tunable knockdown libraries.