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pENTRdeltaGFPmirGE-DroshaT1.5.5
(Plasmid #61084)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 61084 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pENTRL1L2
  • Backbone size w/o insert (bp) 2718
  • Total vector size (bp) 3350
  • Modifications to backbone
    A triple stop codon is inserted at the beginning of GFP ORF to reduce GFP translation. GFP is thus used as a spacer before the miRNAS hairpins. This plasmid contains 3 tandem hairpins that can be excised by BamHI and XbaI to insert your pet mirGE hairpin as described in our article (MTNA, 2014).
  • Vector type
    RNAi

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    deltaGFP
  • Alt name
    3 stop codons
  • Alt name
    no GFP translation
  • gRNA/shRNA sequence
    miRNA (mirGE) triple
  • Species
    H. sapiens (human)

Resource Information

Depositor Comments

From the depositor, regarding a "c" missing from the miRNA: "The mutation is likely from the PCR since the hairpin is amplified by PCR.
I would not use this clone to KD Drosha but in our case, the insert is
used as an indicator of cutting when you use BamHI and XbaI to clone
your own miRNA."

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:

    pENTRdeltaGFPmirGE-DroshaT1.5.5 was a gift from Patrick Salmon (Addgene plasmid # 61084 ; http://n2t.net/addgene:61084 ; RRID:Addgene_61084)
  • For your References section:

    Optimization of Critical Hairpin Features Allows miRNA-based Gene Knockdown Upon Single-copy Transduction. Myburgh R, Cherpin O, Schlaepfer E, Rehrauer H, Speck RF, Krause KH, Salmon P. Mol Ther Nucleic Acids. 2014 Oct 28;3:e207. doi: 10.1038/mtna.2014.58. 10.1038/mtna.2014.58 PubMed 25350582