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Li Human UBDUB CRISPR Knockout Library
(Pooled Library #171531)

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  • Purpose

    Lenti-human-UBDUB-gRNA-puromycin-IRES-CFP is a pooled gRNA library containing 9,274 individual gRNAs against ~1,500 human ubiquitination and de-ubiquitination complex genes (UBDUB). These were constructed in a lentiviral backbone to allow researchers to conduct comprehensive loss-of-function screens in different contexts.

  • Vector Backbone

    LentiGuide-puromycin-IRES-CFP (DNA, 252 KB) was prepared by inserting an IRES-CFP cassette into LentiGuide-Puro. This vector does not express Cas9.

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 171531 Lenti-human-UBDUB-gRNA-puromycin-IRES-CFP pooled library† 1 $430 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. Any SpCas9-expressing plasmid can be used with this library.

Library Details

  • Species
    Human
  • Genes targeted
    1,500
  • gRNAs
    9,274 individual gRNAs
  • Controls
    1,000 non-targeting gRNAs
  • 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
    ∼20 µL
  • Concentration
    50 ng/µL

Resource Information

Depositor Comments

Schematic showing the use of the Li Human UBDUB CRISPR Knockout Library. The pooled library contains 1,000 sgRNAs targeting human ubiquitin ligase genes, 546 sgRNAs targeting epigenetic related genes with an average of 6 sgRNAs per gene, and 100 negative control sgRNAs. An arrow represents library vectors being packaged into lentivirus. The resulting lentiviral pool is used to infect a reporter cell line expressing Cas9. The creation of this cell line is shown in parallel with two steps represented by arrows. First, a reporter cell line is created by knocking in a reporter gene. Second, this cell line is infected with lentivirus containing Cas9 expressing vectors. After infection of reporter cells with the sgRNA pooled library, a new arrow represents FACS sorting of the resulting cell community. A final arrow points towards the last steps of the process which are collection of genomic DNA, recovery of gRNA library, deep sequencing, and identification of enriched gRNAs.
Figure 1: Chunliang Li lab UBDUB CRISPR knockout library

The depositing lab recommends using MAGeCK/MAGeCK-VISPR algorithms (Link opens in a new window) and DESeq2 (Link opens in a new window) for analysis.

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 UBDUB CRISPR knockout library was a gift from Chunliang Li (Addgene #171531)

  • For your References section:

    Interrogating bromodomain inhibitor resistance in KMT2A-rearranged leukemia through combinatorial CRISPR screens. Wright S, Hu J, Wang H, Hyle J, Zhang Y, Du G, Konopleva MY, Kornblau SM, Djekidel MN, Rosikiewicz W, Xu B, Lu R, Yang JJ, Li C. Proc Natl Acad Sci U S A. 2023 Apr 18;120(16):e2220134120. doi: 10.1073/pnas.2220134120. Epub 2023 Apr 10. PMID 37036970.
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