Purposenon-standard AAV2 rep-AAV9.452sub.LUNG1 cap plasmid with AAV cap expression controlled by a tTA-TRE amplification system for mouse lung transduction after intravenous injection
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||184592||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) 8522
Vector typeMammalian Expression, AAV
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Growth Strain(s)NEB Stable
Gene/Insert nameSynthetic construct isolate AAV9.452sub.LUNG1 VP1 gene
Alt nameAAV9.452sub.LUNG1 Cap
Insert Size (bp)2211
Mutation7 amino acid substitution between VP1 452 and VP1 458 from NGSGQNQ to KDNTPGR
- 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 We find that this system increases AAV titers 1.5-5 fold (Ben Deverman, Bryan Simpson and Paul Patterson, unpublished data).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-AAV9.452sub.LUNG1 was a gift from Viviana Gradinaru (Addgene plasmid # 184592 ; http://n2t.net/addgene:184592 ; RRID:Addgene_184592)
For your References section:Targeting the lung epithelium after intravenous delivery by directed evolution of underexplored sites on the AAV capsid. Goertsen D, Goeden N, Flytzanis NC, Gradinaru V. Mol Ther Methods Clin Dev. 2022 Aug 1;26:331-342. doi: 10.1016/j.omtm.2022.07.010. eCollection 2022 Sep 8. 10.1016/j.omtm.2022.07.010 PubMed 35990749