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viral-icon-01.png Viral Service: University of Florida Serotype Testing Panel for the Eye and Brain


As part of our Viral Service, Addgene is producing ready-to-use viral preparations from the University of Florida for enhanced transduction of the eye and other tissues. As part of our standard viral production, these viral vectors undergo quality control, including AAV titration by ddPCR, in vitro and (when possible) in vivo testing, and full sequencing of the final viral vector preparation. These viral vector preparations were produced with the pTR-UF11 plasmid (Addgene #157970).

Viral preparations are formulated in one of the following three buffers. All buffers are supplemented with 0.001% pluronic F68 or 0.014% Tween 20.

  • Phosphate Buffered Saline (PBS)
    • A universal buffer suitable for any application
  • Balanced Salt Solution (BSS)
    • An ophthalmic solution suitable for any application
  • TMN200 (200mM sodium chloride, 1mM magnesium chloride, 20mM Tris at pH 8.0)
    • Suitable for in vitro as well as in vivo delivery where the vector is diluted by the physiologic body fluids, such as blood, vitreous, CSF, etc.
    • Not suitable as a buffer for vectors delivered directly to the brain parenchyma or subretinal space

Serotypes

The following serotypes were developed in Arun Srivastava’s or Shannon Boye’s lab at the University of Florida. References for each serotype can be found below under Citations.

AAV2(Y444F)

The AAV2(Y444F) serotype is demonstrated to have enhanced transduction of the retina following intravitreal injection in mice and marmosets as compared to the AAV2 serotype. The AAV2(Y444F) serotype also displays enhanced transduction of the trabecular meshwork following intracameral injection in mice, rats, and dogs. This serotype was derived from AAV2 capsid and developed by Arun Srivastava’s lab.

AAV2(trpYF)

The AAV2(trpYF) serotype shows enhanced transduction of the retina following intravitreal injection in mice and macaque. This serotype also demonstrates enhanced transduction of cells in culture for ex-vivo conditioning. The AAV2(trpYF) serotype was developed by Arun Srivastava’s lab. It is derived from the AAV2 capsid and contains the following mutations: Y444F, Y500F, and Y730F.

AAV2(Y-F+T-V)

The AAV2(Y-F+T-V) serotype displays enhanced transduction of the retina and is an effective serotype for gene replacement therapy in retinal bipolar cells following intravitreal injection. The AAV2(Y-F+T-V) serotype was developed by Shannon Boye’s lab. It is derived from AAV2 and has the following mutations: Y272F, Y444F, Y500F, Y730F, and T491V.

AAV2(4pMut)dHS

The AAV2(4pMut)dHS serotype displays enhanced transduction of the retina following subretinal administration and enhanced transduction and spread following injection in the brain. The AAV2(4pMut)dHS serotype was developed by Shannon Boye’s lab. It is derived from AAV2 and has the following mutations: Y444F, Y500F, Y730F, T491V, R487G, R585S, and R588T.

AAV6(dbY-F+T-V)

The AAV6(dbY-F+T-V) serotype displays enhanced transduction of hematopoietic stem cells and microglia. The AAV6(dbY-F+T-V) serotype was developed by Arun Srivastava’s lab. It is derived from the AAV6 capsid and contains the following mutations: Y705F, Y731F and T492V.

AAV44.9

The AAV44.9 serotype displays highly efficient transduction and lateral spread in the retina following subretinal injection. It was identified by John Chiorini and characterized by Shannon Boye’s lab.

AAV44.9(E531D)

The AAV44.9(E531D) serotype has increased transduction and lateral spread in the retina following subretinal injection as compared to the parental AAV44.9 serotype. It was developed by Shannon Boye’s lab.

Plasmid Information

The plasmid below is available in each of the above serotypes and buffers. Please click on the plasmid name for ordering information.

ID Name Promoter Description Category PI
157970 pTR-UF11 chimeric CMV/Chicken Beta actin (CBA) GFP Control Sergei Zolotukhin

Citation Information

AAV2(Y444F)

When using the AAV2(Y444F) serotype in future publications, please acknowledge Arun Srivastava and cite Petrs-Silva, et al. 2009. High-efficiency transduction of the mouse retina by tyrosine-mutant AAV serotype vectors. Mol Ther. 2009 Mar;17(3):463-71. PMID: 19066593

Other citations include:

  • Bogner, et al. 2015. Capsid Mutated Adeno-Associated Virus Delivered to the Anterior Chamber Results in Efficient Transduction of Trabecular Meshwork in Mouse and Rat. PLoS One. Jun 8;10(6). PMID: 26052939
  • Lu, et al. 2016. AAV-mediated transduction and targeting of retinal bipolar cells with improved mGluR6 promoters in rodents and primates. Mol Ther. 2016 Aug;23(8-9):680-9. PMID: 27115727
  • Rodriguez-Estevez, et al. 2020. Transduction optimization of AAV vectors for human gene therapy of glaucoma and their reversed cell entry characteristics. Gene Ther. 2020 Apr;27(3-4):127-142. PMID: 31611639

AAV2(trpYF)

When using the AAV2(trpYF) serotype in future publications, please acknowledge Arun Srivastava and cite Petrs-Silva, et al. 2011. Novel properties of tyrosine-mutant AAV2 vectors in the mouse retina. Mol Ther. Feb;19(2):293-301. PMID: 21045809

Other citations include:

  • Chen, et al. 2014. Reprogramming adipose tissue-derived mesenchymal stem cells into pluripotent stem cells by a mutant adeno-associated viral vector. Hum Gene Ther Methods. Feb;25(1):72-82. PMID: 24191859
  • Ye, et al. 2015. Safety and Biodistribution Evaluation in Cynomolgus Macaques of rAAV2tYF-CB-hRS1, a Recombinant Adeno-Associated Virus Vector Expressing Retinoschisin. Hum Gene Ther Clin Dev. Sep;26(3):165-76. PMID: 26390090

AAV2(Y-F+T-V)

When using the AAV2(Y-F+T-V) serotype in future publications, please acknowledge Shannon Boye and cite Kay, et al. 2013. Targeting photoreceptors via intravitreal delivery using novel, capsid-mutated AAV vectors. PLoS One. Apr 26;8(4):e62097. PMID: 23637972

Other citations include:

  • Scalabrino, et al. 2015. Intravitreal delivery of a novel AAV vector targets ON bipolar cells and restores visual function in a mouse model of complete congenital stationary night blindness. Hum Mol Genet. Nov 1;24(21):6229-39. PMID: 26310623

AAV2(4pMut)dHS

When using the AAV2(4pMut)dHS serotype in future publications, please acknowledge Shannon Boye and cite Boye, et al. 2016. Impact of Heparan Sulfate Binding on Transduction of Retina by Recombinant Adeno-Associated Virus Vectors. J Virol. Mar 28;90(8):4215-4231. PMID: 26865709

Other citations include:

  • Kanaan, et al. 2017. Rationally Engineered AAV Capsids Improve Transduction and Volumetric Spread in the CNS. Mol Ther Nucleic Acids. Sep 15;8:184-197. PMID: 28918020

AAV6(dbY-F+T-V)

When using the AAV6(dbY-F+T-V) serotype in future publications, please acknowledge Arun Srivastava and cite Ling, et al. 2016. High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors: Strategies for Overcoming Donor-Variation and Implications in Genome Editing. Sci Rep. Oct 19;6:35495. PMID: 27759036

Other citations include:

  • Ling, et al. 2016. High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors: Strategies for Overcoming Donor-Variation and Implications in Genome Editing. Sci Rep. Oct 19;6:35495. PMID: 27759036
  • Rosario, et al. 2016. Microglia-specific targeting by novel capsid-modified AAV6 vectors. Mol Ther Methods Clin Dev. Apr 13;3:16026. PMID: 27308302

AAV44.9

When using the AAV44.9 serotype in future publications, please acknowledge John Chiorini and Shannon Boye and cite Boye, et al. 2020. Novel AAV44.9-Based Vectors Display Exceptional Characteristics for Retinal Gene Therapy. Mol Ther. Jun 3;28(6):1464-1478. PMID: 32304666

AAV44.9(E531D)

When using the AAV44.9(E531D) serotype in future publications, please acknowledge Shannon Boye and cite Boye, et al. 2020. Novel AAV44.9-Based Vectors Display Exceptional Characteristics for Retinal Gene Therapy. Mol Ther. Jun 3;28(6):1464-1478. PMID: 32304666

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