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Turner Lab Human messenger-RBP sgRNA Library
(Pooled Library #168791)

  • Purpose

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

  • Vector Backbone
    • pKLV2_hU6_BbsI_ccdB_BbsI_iScaffold_mPGK_puro_T2A_Thy1.1
      • Does not express Cas
      • Not available through Addgene
    • Plasmid annotation and sequence (Genbank) (12.7 KB)
    • Plasmid description: PKLV2_hU6_BbsI-ccdB-BbsI_iScaffold_mPGK_puro-2A-Thy1.1 (PKLV2_sgRNA_puro-Thy1.1) was generated in two rounds of cloning from Addgene Plasmid 67974: pKLV2-U6gRNA5(BbsI)-PGKpuro2ABFP-W. Firstly, a PCR product encoding Thy1.1 and the appropriate flanking sequences was generated and ligated (Gibson assembly) into a XhoI + NotI linearised vector (Addgene Plasmid 67974). Secondly, a PCR product encoding the ccdB toxic gene, a region validated to contain the ccdB gene’s uncharacterised promoter, and the appropriate flanking sequences were generated and then ligated (Gibson assembly) into a BbsI linearised intermediate vector (PKLV2_hU6_BbsI-BbsI_iScaffold_mPGK_puro-2A-Thy1.1).

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 168791 Human 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).

Library Details

  • Species
    Human
  • Genes targeted
    725
  • gRNAs
    8,260
  • Controls
    500 non-targeting sgRNAs
  • Lentiviral Generation
    3rd

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
    ∼20µL
  • Concentration
    50ng/µL
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:

    Human messenger-RBP sgRNA Library was a gift from Martin Turner (Addgene #168791)
  • For your References section:

    RNA Binding Proteins As Regulators of Oxidative Stress Identified by a Targeted CRISPR-Cas9 Single Guide RNA Library. Turner DJ, Turner M CRISPR J. 2021 Jun;4(3):427-437. doi: 10.1089/crispr.2020.0116. Epub 2021 Jun 4. PubMed 34096786