Pooled Libraries
Mouse messenger-RBP sgRNA Library

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 Cas
  - Not available through Addgene
- Plasmid annotation and sequence (Genbank) (12.5 KB)
- Plasmid description: MSCV_hU6_BbsI-ccdB-BbsI_iScaffold_mPGK_puro-2A-Thy1.1 (MSCV_sgRNA_puro-Thy1.1) was generated in one round of cloning from Addgene Plasmid 27490: MIGR1. A PCR product encoding hU6_BbsI-ccdB-BbsI-iScaffold_mPGK_Puro-2A-Thy1.1 was ligated by Gibson assembly into a XhoI + SalI linearised vector.

Depositing Labs

Martin Turner
Publication

Turner et al. eLife. 2022 Apr 22;11:e72313. doi: 10.7554/eLife.72313. (How to cite )

Ordering

Item	Catalog #	Description	Quantity	Price (USD)
Pooled Library	169082	Mouse messenger-RBP sgRNA Library^†	1		$400	Add to Cart

Available to Academic and Nonprofits Only

^† A Cas9 plasmid is NOT included with this item and will have to be ordered separately. Can be used in conjunction with lentiCas9-Blast (Addgene #52962) or established Cas9 mouse models.

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
- UBMTA

Depositor Comments

Turner Lab Mouse messenger-RBP Library Overview

**(A)** Schematic representation of retrovirus sgRNA library that encodes puromycin resistance and CD90.1 (Thy1.1) selection markers, and encodes an improved sgRNA scaffold that lacks four consecutive thymine residues and has an extended duplex region. (Top) Schematic representation of vector backbone that encodes ccdB toxin gene to improve cloning efficiency. **(B)** Distribution of the representation of sgRNAs in library. The library is normally distributed and tightly distributed.

Dang, Y. et al. Optimizing sgRNA structure to improve CRISPR-Cas9 knockout efficiency. Genome Biol. (2015) doi:10.1186/s13059-015-0846-3.

Please visit https://www.biorxiv.org/content/10.1101/2021.07.21.453193v1 (Link opens in a new window) for bioRxiv preprint.

How to cite this pooled library ( Back to top )

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:

Mouse messenger-RBP sgRNA Library was a gift from Martin Turner (Addgene #169082)
For your References section:

A functional screen of RNA binding proteins identifies genes that promote or limit the accumulation of CD138+ plasma cells. Turner DJ, Saveliev A, Salerno F, Matheson LS, Screen M, Lawson H, Wotherspoon D, Kranc KR, Turner M. eLife 2022;11:e72313. doi: 10.7554/eLife.72313 PMID: 35451955

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