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CHyMErA Knockout Libraries
(Pooled Library #155199, #155200)

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  • Purpose

    The Cas Hybrid for Multiplexed Editing and screening Applications (CHyMErA) lentiviral libraries express hybrid Cas9-Cas12a guide RNAs under a single U6 promoter.

    The Paralog & Dual-targeting hybrid guide RNA Library targets human paralog pairs with single- and combinatorial-targeting hgRNAs and can be used to identify genetic interactions between paralogs. The library also contains single- and dual-targeting hybrid guides that target ~5,000 highly expressed genes permitting capture of gene ablation phenotypes with increased sensitivity.

    The Exon-deletion hgRNA Library deletes human alternative cassette exons and disrupts genes containing alternative exons, allowing investigation into alternative exon function.

  • Vector Backbone

    pLCHKO (Plasmid #155048) - does not express Cas9 or Cas12a

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 155199 CHyMErA Paralog & Dual-targeting hgRNA pooled library† 1 $540 Add to Cart
Pooled Library 155200 CHyMErA Exon-deletion hgRNA pooled library† 1 $540 Add to Cart
Available to Academic and Nonprofits Only

  • † SpCas9 and LbCas12a plasmids are NOT included with this item and will have to be ordered separately. Can be used in conjunction with lenti-Cas9-2A-BLAST (Addgene #73310) and plenti-Lb-Cas12a-2xNLS (Addgene #155046), or with a cell line expressing both SpCas9 and LbCas12a.

Library Details

  • Species
    Human
  • Genes targeted


    Paralog & Dual-targeting library: 6,333 genes (1,344 paralogs & 4,993 genes)
    Exon-deletion library: 2,157 alternative cassette exons in 1,745 genes

  • hgRNAs


    Paralog & Dual-targeting library: 92,746
    Exon-deletion library: 91,948

  • Controls


    Paralog & Dual-targeting library: 4,993 intergenic and non-targeting hgRNAs
    Exon-deletion library: 1,647 intergenic control hgRNAs

  • 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


Screening analysis is described in:
Pooled Lentiviral CRISPR-Cas9 Screens for Functional Genomics in Mammalian Cells. Aregger M, Chandrashekhar M, Tong AHY, Chan K, Moffat J. Methods Mol Biol. 2019;1869:169-188. doi: 10.1007/978-1-4939-8805-1_15. PubMed 30324523 (Link opens in a new window)

The depositing lab has provided analysis algorithms and R code in the following Github repositories:

CHyMERa Dual-targeting Scoring (Link opens in a new window)
CHyMERa Exon-deletion Scoring (Link opens in a new window)

Read depositor Jason Moffat's description of CHyMErA in this Research Communities "Behind The Paper" article (Link opens in a new window).

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:

    CHyMErA Paralog & Dual-targeting hgRNA Pooled Library was a gift from Jason Moffat and Benjamin Blencowe (Addgene #155199)
    CHyMErA Exon-deletion hgRNA Pooled Library was a gift from Jason Moffat and Benjamin Blencowe (Addgene #155200)

  • For your References section:

    Genetic interaction mapping and exon-resolution functional genomics with a hybrid Cas9-Cas12a platform. Gonatopoulos-Pournatzis T, Aregger M, Brown KR, Farhangmehr S, Braunschweig U, Ward HN, Ha KCH, Weiss A, Billmann M, Durbic T, Myers CL, Blencowe BJ, Moffat J. Nat Biotechnol. 2020 May;38(5):638-648. doi: 10.1038/s41587-020-0437-z. PubMed 32249828 (Link opens in a new window)
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