Turner Lab Mouse messenger-RBP sgRNA Library
(Pooled Library #169082)

• Purpose:

  This retroviral CRISPR knockout library targets mouse messenger RNA-binding proteins with 10 single guide RNAs (sgRNAs) per gene. The retroviral backbone encodes both puromycin resistance and the CD90.1 selection marker.

• Vector Backbone:

  MSCV_hU6_BbsI-ccdB-BbsI_iScaffold_mPGK_puro-2A-Thy1.1 - does not express Cas9

• Depositing Labs: Martin Turner
• Publication: Turner et al. eLife. 2022 Apr 22;11:e72313. doi: 10.7554/eLife.72313.

Library Details

• Species: Mouse
• Genes targeted: 1,300
• gRNAs: 13,500
• Controls: 500 non-targeting sgRNAs

Library Shipping

This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.

• Volume: ∼20 µL
• Concentration: 50 ng/µL

Resource Information

• Protocols:
  • Library Transformation and Sequencing Protocols (20.4 KB)
• • NGS Primer Table (9.4 KB)
  • Table of gene targets, gRNA sequences, and count data (392.2 KB)

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA

Depositor Comments

Figure 1: Mouse messenger-RBP Library Overview. A) Schematic representation of vector backbone and retrovirus sgRNA library components. The vector backbone encodes ccdB toxin gene for cloning (replaced by sgRNA seed sequence), an improved sgRNA scaffold that lacks four consecutive thymine bases and has an extended duplex region, and puromycin resistance and CD90.1 (Thy1.1) selection markers. B) Distribution of the representation of sgRNAs in the library is normally distributed and tightly distributed.

Please visit https://doi.org/10.1101/2021.07.21.453193 for bioRxiv preprint.

See also Dang, Y. et al. Optimizing sgRNA structure to improve CRISPR-Cas9 knockout efficiency. Genome Biol. 2015 Dec 15:16:280. doi: 10.1186/s13059-015-0846-3 .