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Human Genome-wide Intron Tagging Libraries
(Pooled Library #216133, #216134)

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  • Purpose

    Library of intron-targeting sgRNAs for pooled tagging of proteins with GFP or other protein tags at their endogenous genomic locus using an intron tagging approach. When used with other intron tagging plasmids, the intron-targeting sgRNAs specify where a synthetic exon is placed in the genome.

    The library is not used for CRISPR knockout, activation, or inhibition, but for pooled protein tagging using a CRISPR/Cas9 based intron tagging approach.

  • Vector Backbone

    Human Genome-wide Intron Tagging Library, Frame 0: CROPseq-Guide-Puro (Plasmid #86708) - does not express Cas9

    Human Genome-wide Intron Tagging Library, Frame 1: CROPseq-Guide-BSD (Plasmid #216123) - does not express Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 216133 Human Genome-wide Intron Tagging Library, Frame 0† 1 $540 Add to Cart
Pooled Library 216134 Human Genome-wide Intron Tagging Library, Frame 1† 1 $540 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. For pooled protein tagging using an intron tagging approach, the library can be used with any regular Cas9 plasmid and a DNA donor plasmid or minicircle. For example Intron-Tagging-pX330-Cas9-mCherry (Addgene #159742) and Intron-Tagging-EGFP-Donor (Addgene #159740) or Intron-Tagging-EGFP-Frame0-Parental-Minicircle (Addgene #216124).

Library Details

  • Species
    Human
  • gRNAs

    1–3 sgRNAs per intron

    • Frame 0 library: 90,657 sgRNAs targeting 73,817 introns of 14,158 genes
    • Frame 1 library: 72,580 sgRNAs targeting 51,939 introns of 14,011 genes.
  • Controls
    1,000 non-targeting sgRNAs per library
  • 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
    ∼25 µL
  • Concentration
    50 ng/µL

Resource Information

Depositor Comments

The left panel (frame 0) has an arrow pointing down from a “minicircle DNA donor for frame 0 introns” with “Cas9 cut sites” leading to a GFP gene flanked by a “splice acceptor” and a “splice donor”. To the left is a drawing of some unlabeled cells and to the right is a drawing of a “GFP-tagged cell pool” with cells showing GFP in different patterns throughout the cells (punctate or homogenous in the membrane, cytosol, nucleus, etc). This leads to a linear piece of DNA containing Cas9 cut sites. An arrow from below leads to this same DNA with the “Human Genome-wide Intron Tagging Library, Frame 0” drawn as virus particles with different cargos inside. Asp (AAG) and Lys (AAG) on the left and Ala (GCT) and Leu (CTC) on the right create the Frame 0 intron. The second panel is identical except mScarlet takes the place of GFP and Thr (ACA) and G on the left and ly (GG) and Val (GTG) on the right create the Frame 1 intron. The third panel below is labeled “2 rounds of pooled intron tagging for multicolor cell pools.” The left side of this panel is identical to the top left. The right side of the bottom panel is identical to the top right except the unlabeled cells on the left are gone (“GFP-tagged cell pool takes their place) and the cells on the right side are a “double-tagged cell pool” showing GFP and mScarlet in different patterns throughout the cells.

Figure 1: Overview of pooled intron tagging, including tagging for multicolor cell pools.

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 Genome-wide Intron Tagging Library, Frame 0 were a gift from Stefan Kubicek (Addgene #216133)
    Human Genome-wide Intron Tagging Library, Frame 1 were a gift from Stefan Kubicek (Addgene #216134)

  • For your References section:

    Pooled multicolour tagging for visualizing subcellular protein dynamics. Reicher A, Reiniš J, Ciobanu M, Růžička P, Malik M, Siklos M, Kartysh V, Tomek T, Koren A, Rendeiro AF, Kubicek S. Nat Cell Biol. 2024 May;26(5):745-756. doi: 10.1038/s41556-024-01407-w. PubMed 38641660 (Link opens in a new window)
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