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pMSCV-Cas9-2A-GFP-sgRNA
(Plasmid #124889)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 124889 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pMCSV
  • Backbone size w/o insert (bp) 3000
  • Total vector size (bp) 10500
  • Vector type
    Retroviral

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Cas9
  • Alt name
    S. pyogenes CRISPR-Cas9
  • Species
    Synthetic; Streptococcus Pyogenes
  • Insert Size (bp)
    4200
  • Promoter MSCV
  • Tag / Fusion Protein
    • 2A-GFP (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site XhoI (not destroyed)
  • 3′ cloning site EcoRI (not destroyed)
  • 5′ sequencing primer Gag
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Guide RNA to be cloned into BbsI site. Supplied modified golden-gate cloning method is required given that extra BbsI site is located within the 2A sequence.

To design guide RNAs:
-add ACCG to 5' end of positive strand (positive strand defined as strand which will contact PAM sequence at 3' end)
-add AAAC to 5' end of negative strand

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:

    pMSCV-Cas9-2A-GFP-sgRNA was a gift from Harvey Lodish (Addgene plasmid # 124889 ; http://n2t.net/addgene:124889 ; RRID:Addgene_124889)
  • For your References section:

    Efficient CRISPR-Cas9 mediated gene disruption in primary erythroid progenitor cells. Li H, Shi J, Huang NJ, Pishesha N, Natarajan A, Eng JC, Lodish HF. Haematologica. 2016 Jun;101(6):e216-9. doi: 10.3324/haematol.2015.135723. Epub 2016 Mar 11. 10.3324/haematol.2015.135723 PubMed 26969085