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Watermelon Library
(Pooled Library #155257)

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  • Purpose

    The Watermelon library is a high complexity lentiviral barcode library for simultaneous tracing of clonal lineages as well as the transcriptional and proliferative state of each cell in the population. Lineage tracing is achieved by mapping a clone-specific expressed DNA barcode in the 3′ untranslated region of an mNeonGreen protein, and proliferation history is monitored by the dilution of a doxycycline (dox)-inducible H2B-mCherry transgene.

  • Vector Backbone

    Watermelon backbone (Plasmid #155258)

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 155257 Watermelon Library 1 $640 Add to Cart
Available to Academic and Nonprofits Only

Library Details

  • Number of Barcodes
    5 x 106
  • 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/uL

Resource Information

Depositor Comments

Four schematics describing the Watermelon system. The Watermelon construct consists of semi-random barcode, NLS-mNeon, and H2B-mCherry (inducible). The lentiviral Watermelon library is used to infect cells, the infected cells then have unique barcodes, and with the addition of doxycycline the cells reach their transcriptional and proliferative cell state. After removal of doxycycline, FACS sorting can track proliferation. And lineage tracing uses scRNA-Seq to track individual clones.

Figure 1. The Watermelon system for simultaneous tracing of clonal lineages and transcriptional and proliferative state of individual cells.

The data and code related to this library is available on GitHub (Link opens in a new window).

The file "Watermelon Library Sequence (Primers marked)" linked above provides an example of the plasmid sequence. Please see the Nature publication section "Cloning of the Watermelon library" to see how the final sequence compares to the Watermelon backbone (Plasmid #155258).

NOTE: Lentiviral vectors can recombine during library amplification, resulting in a ~6 kb and ~3.5 kb band. The recombined backbone product should not adversely affect library function as it does not contain lentiviral packaging sequences. If these additional bands represent a large percentage of the product after amplification, we recommend starting with a larger prep (gigaprep) from the original sample or gel purifying the amplified library to remove the recombinant band and performing another transformation with the purified sample.

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:

    Watermelon library was a gift from Aviv Regev and Joan Brugge (Addgene #155257)

  • For your References section:

    Cycling cancer persister cells arise from lineages with distinct programs. Oren Y, Tsabar M, Cuoco MS, Amir-Zilberstein L, Cabanos HF, Hutter JC, Hu B, Thakore PI, Tabaka M, Fulco CP, Colgan W, Cuevas BM, Hurvitz SA, Slamon DJ, Deik A, Pierce KA, Clish C, Hata AN, Zaganjor E, Lahav G, Politi K, Brugge JS, Regev A. Nature. 2021 Aug 11. pii: 10.1038/s41586-021-03796-6. doi: 10.1038/s41586-021-03796-6. PubMed 34381210
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