Purposenon-standard AAV2 rep-AAV.CAP-B10 cap plasmid with AAV cap expression controlled by a tTA-TRE amplifcation system
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||175004||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 6311
- Total vector size (bp) 8543
Vector typeMammalian Expression, AAV
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Growth Strain(s)NEB Stable
Gene/Insert nameSynthetic construct isolate AAV.CAP-B10 VP1 gene
Alt nameAAV.CAP-B10 Cap
Insert Size (bp)2232
Mutation7 amino acid substitution between VP1 452 and VP1 460 from NGSGQNQ to DGAATKN
- Promoter p41
- Cloning method Gibson Cloning
- 5′ sequencing primer unknown (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
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 We find that this system increases AAV titers 1.5-5 fold (Ben Deverman, Bryan Simpson and Paul Patterson, unpublished data).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.
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-B10 was a gift from Viviana Gradinaru (Addgene plasmid # 175004 ; http://n2t.net/addgene:175004 ; RRID:Addgene_175004)
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