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Addgene

pUCmini-iCAP-AAV.CAP-B22
(Plasmid #175005)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 175005 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pUC57-mini
  • Backbone size w/o insert (bp) 6311
  • Total vector size (bp) 8543
  • Vector type
    Mammalian Expression, AAV

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    Synthetic construct isolate AAV.CAP-B22 VP1 gene
  • Alt name
    AAV.CAP-B22 Cap
  • Species
    Synthetic
  • Insert Size (bp)
    2232
  • Mutation
    7 amino acid substitution between VP1 452 and VP1 460 from NGSGQNQ to DGQSSKS
  • Promoter p41

Cloning Information

Resource Information

Terms and Licenses

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

Depositor Comments

This plasmid is based on the pUCmini-iCAP-PHP.eB plasmid. Please note that this is a non-standard rep-cap construct. It uses a tTA-TRE amplification loop to increase Capsid expression and AAV production. We find that this system increases AAV titers 1.5-5 fold (Ben Deverman, Bryan Simpson and Paul Patterson, unpublished data). This is a tet-off system, so no dox or tet is needed to turn it on. It can be used like any other rep-cap plasmid. This system should only present a problem if the rAAV genome to be packaged has a tet responsive element that directs expression of a protein that affected the health of the production cells. This work supported by NIH BRAIN Initiative grant UF1MH128336 . The introduction of an XbaI restriction site for ease of cloning introduces a K449R mutation, which does not have an overt effect on vector production or transduction.

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:

    pUCmini-iCAP-AAV.CAP-B22 was a gift from Viviana Gradinaru (Addgene plasmid # 175005 ; http://n2t.net/addgene:175005 ; RRID:Addgene_175005)
  • For your References section:

    AAV capsid variants with brain-wide transgene expression and decreased liver targeting after intravenous delivery in mouse and marmoset. Goertsen D, Flytzanis NC, Goeden N, Chuapoco MR, Cummins A, Chen Y, Fan Y, Zhang Q, Sharma J, Duan Y, Wang L, Feng G, Chen Y, Ip NY, Pickel J, Gradinaru V. Nat Neurosci. 2022 Jan;25(1):106-115. doi: 10.1038/s41593-021-00969-4. Epub 2021 Dec 9. 10.1038/s41593-021-00969-4 PubMed 34887588