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pRUSH
(Plasmid #54479)

Full plasmid sequence is not available for this item.

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 54479 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    ROSA26-1
  • Backbone manufacturer
    Phil Soriano, Addgene Plasmid #21714
  • Modifications to backbone
    Modified by deleting the HpaI site, converting the XhoI site to AscI, and cloning a splice acceptor-GFP-polyA into the XbaI site. Cre-lox regulated U6 cassette derived from pSICOR, Addgene plasmid 11579, (Ventura et al., 2004) was modified by replacing the lentiviral GFP gene with drug selection markers (pgk-neo), and cloned into the XbaI site 3' of GFP. Unique HpaI and XhoI sites were maintained for single step short hairpin oligonucleotide cloning.
  • Vector type
    Mammalian Expression, Mouse Targeting, RNAi, Cre/Lox
  • Promoter U6
  • Selectable markers
    Neomycin (select with G418)
  • Tags / Fusion Proteins
    • SA-EGFP-polyA (N terminal on backbone)
    • loxP (N terminal on backbone)
    • U6 promoter (N terminal on backbone)
    • pgk-neo (C terminal on backbone)
    • loxP (C terminal on backbone)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ sequencing primer RUSH-for CACAAAAGGAAACTCAC CCTAA
  • 3′ sequencing primer RUSH-rev CTAAAGCGCATGCTCCA GAC
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Unique HpaI and XhoI sites were maintained for single step short hairpin oligonucleotide cloning.

Using the primers described above yields 231bp and 177bp for positive and negative products, respectively.

How to cite this plasmid ( Back to top)

These plasmids 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:

    pRUSH was a gift from Laurie Jackson-Grusby (Addgene plasmid # 54479 ; http://n2t.net/addgene:54479 ; RRID:Addgene_54479)
  • For your References section:

    RUSH and CRUSH: a rapid and conditional RNA interference method in mice. Brown JR, Zetsche B, Jackson-Grusby L. Genesis. 2014 Jan;52(1):39-48. doi: 10.1002/dvg.22718. Epub 2013 Oct 26. 10.1002/dvg.22718 PubMed 24166816