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BARBEKO sgRNA Library
(Pooled Library #174163)

  • Purpose

    BARBEKO, short for iBARed cytosine Base Editing-mediated gene KnockOut, leverages CRISPR cytosine base editors for genome-scale knockout screens in human cells by perturbing gene start codons, splice sites, or by introducing premature termination codons. It is integrated with iBAR, a design in the sgRNA scaffold for improving screening quality and efficiency.

    Each sgRNA construct contains one iBAR sequence and each sgRNA is followed by four iBARs (214,008 sgRNAiBARs in total).

  • Vector Backbone

    pLenti-sgRNA-Lib - does not express Cas9

    This library is designed for gene knockout screens by using CRISPR cytosine base editors (CBE), but not Cas9. Library was re-constructed into lentiviral sgRNA-iBAR backbone according to the method in Zhu et al., 2019 PMID: 30678704.

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 174163 BARBEKO sgRNA library 1 $500 Add to Cart
Available to Academic and Nonprofits Only
  • An spCas9-based cytosine base editing plasmid is NOT included with this item and will have to be ordered separately. Can be used in conjunction with pCMV_AncBE4max (Addgene #112094) or pLenti-AncBE4max (Addgene #174164).

Library Details

  • Species
    Human
  • Genes targeted
    17,501
  • gRNAs
    53,502
  • Controls


    Non-targeting controls: 500
    Negative controls targeting safe-harbor regions: 499

  • 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

Depositor Comments

Graphic summary of BARBEKO sgRNA pooled library
  • CBE with sgRNAs targeting start codons, splice acceptor sites, splice donor sites, and codons of glutamine, arginine or tryptophan disrupting gene functions.


The CBE-expressing cassette from pCMV_AncBE4max (Addgene #112094) was transferrred into a lentiviral backbone to create pLenti-AncBE4max (Addgene #174164). To efficiently package the CBE into lentivirus, concentrate the lentiviral supernatants before use.

For cell fitness screens, ZFCiBAR algorithm can be used and obtained from https://github.com/wolfsonliu/zfc. For positive selection screens, MAGeCKiBAR algorithm can be used for analysis (Zhu et al., 2019): https://bitbucket.org/WeiLab/mageck-ibar/.

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:

    BARBEKO sgRNA library was a gift from Wensheng Wei (Addgene #174163)
  • For your References section:

    Genome-wide interrogation of gene functions through base editor screens empowered by barcoded sgRNAs. Xu P, Liu Z, Liu Y, Ma H, Xu Y, Bao Y, Zhu S, Cao Z, Wu Z, Zhou Z, Wei W. Nat Biotechnol. 2021 Jun 21. pii: 10.1038/s41587-021-00944-1. doi: 10.1038/s41587-021-00944-1. PubMed 34155407