Purposenon-standard AAV2 rep-AAV-PHP.eB cap plasmid with AAV cap expression controlled by a tTA-TRE amplifcation system
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||103005||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector typeMammalian Expression, AAV
Growth in Bacteria
Growth Strain(s)NEB Stable
Gene/Insert nameSynthetic construct isolate AAV-PHP.eB VP1 gene
Alt nameAAV-PHP.eB Cap
- Promoter p41
- Cloning method Gibson Cloning
- 5′ sequencing primer unknown (Common Sequencing Primers)
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 We find that this system increases AAV titers 1.5-5 fold (Ben Deverman, Bryan Simpson and Paul Patterson, unpublished data).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-PHP.eB was a gift from Viviana Gradinaru (Addgene plasmid # 103005 ; http://n2t.net/addgene:103005 ; RRID:Addgene_103005)
For your References section:Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V. Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub 2017 Jun 26. 10.1038/nn.4593 PubMed 28671695