pUCmini-iCAP-AAV9.452sub.LUNG1
(Plasmid #184592)

• Purpose: non-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
• Depositing Lab: Viviana Gradinaru
• Publication: Goertsen et al Mol Ther Methods Clin Dev. 2022 Aug 1;26:331-342. doi: 10.1016/j.omtm.2022.07.010. eCollection 2022 Sep 8.

Backbone

• Vector backbone: pUC57-mini
• Backbone size w/o insert (bp): 6311
• Total vector size (bp): 8522
• 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 AAV9.452sub.LUNG1 VP1 gene
• Alt name: AAV9.452sub.LUNG1 Cap
• Species: Synthetic
• Insert Size (bp): 2211
• Mutation: 7 amino acid substitution between VP1 452 and VP1 458 from NGSGQNQ to KDNTPGR
• Promoter: p41

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: unknown

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

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 We find that this system increases AAV titers 1.5-5 fold (Ben Deverman, Bryan Simpson and Paul Patterson, unpublished data).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. 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.