sgRNA libraries for Pseudomonas aeruginosa PA14
(Pooled Library #234855, #1000000254)
-
Purpose
A comprehensive CRISPRi library has been designed for inducible, genome-wide gene repression in Pseudomonas aeruginosa PA14. This library consists of two subsets: Set 1, containing 5,981 sgRNAs targeting distinct genes, and Set 2, comprising 5,971 sgRNAs targeting the same genes as Set 1 but designed for different PAM regions. The sgRNAs in Set 1 and Set 2 complement each other, offering a robust and versatile approach to gene targeting.
The library has been meticulously engineered for efficient transformation into E. coli strains such as S17 or other E. coli strains carrying the lambda pir gene, which is necessary for R6K ori replication. This system enables the construction of a tetracycline-inducible CRISPRi platform, facilitating precise and controlled gene repression studies.
-
Vector Backbone
pCRISPRi-ccdB (Plasmid #226432) - expresses dCas9
-
Depositing Labs
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |||
---|---|---|---|---|---|---|---|
Pooled Library | 234855 | Set 1, one tube, contains sgRNA library for Pseudomonas aeruginosa PA14 | 1 | $380 | Add to Cart | ||
Pooled Library | 1000000254 | Two tubes, each with a half-library (Set 1 and 2)
| 1 | $540 | Add to Cart |
Library Details
-
SpeciesPseudomonas aeruginosa
-
Genes targeted
- Set 1: 5,981
- Set 2: 5,971
-
gRNAs1 sgRNA per gene in each set
Library Shipping
Each library is delivered in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.
-
Volume∼25 µL
-
Concentration100 ng/µL
Resource Information
-
Protocols
-
Depositor Data
-
Scripts
- The 2FAST2Q method (Link opens in a new window)can be used for analysis
-
Terms and Licenses
Academic/Nonprofit TermsIndustry Terms- Not Available to Industry
Trademarks- Zeocin® is an InvivoGen trademark.
Depositor Comments
Set 1 is a primary set of 5,981 sgRNAs with high specificity targeting all operons and genes in the P. aeruginosa PA14 genome. Set 2 is a secondary set of 5,971 sgRNAs providing alternative guides to ensure full genome coverage in P. aeruginosa PA14.
These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
-
For your Materials & Methods section:
sgRNA libraries Set 1 for Pseudomonas aeruginosa PA14 was a gift from Xue Liu (Addgene #234855 ; http://n2t.net/addgene:234855 ; RRID:Addgene_234855)
sgRNA libraries Set 1 and 2 for Pseudomonas aeruginosa PA14 was a gift from Xue Liu (Addgene #1000000254) -
For your References section:
Genome-wide CRISPRi-seq Identified Ferredoxin-NADP Reductase FprB as a Synergistic Target for Gallium Therapy in Pseudomonas aeruginosa. Zhang Y, Zhang T, Xiao X, Kawalek A, Ou J, Ren A, Sun W, de Bakker V, Liu Y, Li Y, Yang L, Ye L, Jia N, Veening JW, Wang Y, Liu X. bioRxiv 2024.09.01.610675. doi: 10.1101/2024.09.01.610675.