URL: http://www.addgene.org/103005
Proper Citation: RRID:Addgene_103005
Insert Name: Synthetic construct isolate AAV-PHP.eB VP1 gene
Organism: Synthetic
Bacterial Resistance: Ampicillin
Defining Citation: PMID:28671695
Vector Backbone Description: Vector Backbone:pUC57-mini; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin
Comments: 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 We find that this system increases AAV titers 1.5-5 fold (Ben Deverman, Bryan Simpson and Paul Patterson, unpublished data).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. 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.
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Source: Addgene