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AAV Capsid Barcode Kit-BBB
AAV Capsid Barcode Kit-BBB
  1. ATHENA-I-Capsid-Library-2.png
  2. AAV Capsid Barcode Kit-BBB
  3. AAV Capsid Barcode Kit-BBB

AAV Capsid Barcode Kit-BBB

Price range: $9,999.00 through $49,999.00

AAV Capsid Barcode Kit-BBB  is designed to systematically evaluate different AAV serotypes or variants of their activity to cross blood-brain-barrier (BBB).  Gene therapy targeting CNS diseases faces significant challenges primarily due to the BBB, which restricts the entry of therapeutic delivery vehicles. AAV9 has become a focal point in CNS gene therapy due to its ability to cross the BBB in mouse models. AAV9 serves as a foundational template for developing new AAV vectors with enhanced BBB penetration capabilities, such as AAV-Php.eB. However, AAV capsids that were modified for BBB crossing in mice have shown limited success in non-human primates (NHPs). Moreover, the species of mouse and NHP also affect the crossing-BBB activity of AAVs. This library includes a combination of 15 common AAV capsids and 9 reported crossing BBB AAV capsids, providing researchers with a comprehensive set of tools for targeted gene delivery to the brain. Researchers can conduct comparative studies to assess the transduction efficiency, specificity, and safety profiles of different AAV capsids targeting to CNS tissues using AAV Capsid Barcode Kit-BBB.

  • SKU:DH005001B
  • AAV Expression Cassette: AAV-CAG-EGFP-WPRE-BCs-bGHpolyA.
  • AAV Capsid List:
  • AAV status: Mixed viruses.
  • AAV Titer: ~1X10^13 VG/ml
  • BC design: (N)12-ACGGAAATACGATGTCGGGA-(N)12
  • Production: One AAV capsid and assigned 3 barcode plasmids were co-transfected to make one specific capsid AAV vector.
  • Purification: Each AAV capsid were produced(HEK 293T), purification(two rounds of CsCl) and titration(qPCR), separately.
  • Storage Buffer: 0.001% F-68/DPBS with additional 150mM NaCl.
  • Storage:  -80°C for long term (> 1 year);  -20°C for short term(1-2 months);  4 °C for 1-2 weeks.
  • Shipping: Dry ice.
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AAV Capsid Barcode Kits-Tissues

Tissue-specific AAV capsid Barcode Kits are designed to systematically evaluate AAV capsid variants that have been engineered to exhibit enhanced tropism for specific target tissues. It also includes 15 commonly used AAV serotypes that serve as controls, which are essential for benchmarking and comparative purposes.

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AAV Capsid Barcode Kits-Common

Common AAV Capsids: AAV1, AAV2, AAV3B, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh.10, AAV11, AAV12, AAV13, AAVrh.74, AAV-DJ, AAV2-Retro

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Design of AAV Capsid Barcode Kits

AAV Capsid Barcode Kits are known AAV capsid selection kits build on our ATHENA I platform. In the ATHENA I, each capsid variant is associated with three different DNA-barcoded genomes, and all three of these genomes carry the same reporter gene. The use of three DNA barcodes for one capsid variant can minimize experimental variations and improve the accuracy of the results in high-throughput screening or selection processes.

In the AAV Capsid Barcode Kits,  AAV genomes carried different  promoters and different reporter genes, such as AAV-CAG-EGFP, AAV-CMV-mCherry, and scAAV-CMV-mCherry. The presence of the reporter gene allows for the enrichment of transgene-expressing cells, making it possible to identify and isolate cells that have taken up and expressed the AAV genomes.

The unique barcodes  are located before Poly signal. NGS technology is used to analyze the barcode data after AAV infection or injection.  Researchers can use the unique DNA barcodes to evaluate the distribution of each capsid variant for different types of target cells. This screening method helps identify which AAV capsids are most effective at transducing specific cell types. In addition to DNA barcode screening, transcribed RNA barcodes can be evaluated to assess the potential of different capsids for each target cell. This approach provides insights into the transcriptional activity of the AAV genomes in the context of specific cell types.

Common AAV Capsid List: AAV1, AAV2, AAV3B, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh.10, AAV11, AAV12, AAV13, AAVrh.74, AAV-DJ, AAV2-Retro

 

ATHENA I Known Capsid Library

 

Example ​1:

AAV Capsid Kit: AAV Capsid Barcode Kit-Common-CAG-EGFP

Animal model: C57B6

Injection Route:  IV injection

Time Point: 2 weeks post injection

Analysis method: NGS for BCs

Results: Fold enrichment compared with input AAV viruses

AAV Capsid Library(I)-Common

References:

  1. In Vivo Selection Yields AAV-B1 Capsid for Central Nervous System and Muscle Gene Therapy. Choudhury SR, Fitzpatrick Z, Harris AF, Maitland SA, Ferreira JS, Zhang Y, Ma S, Sharma RB, Gray-Edwards HL, Johnson JA, Johnson AK, Alonso LC, Punzo C, Wagner KR, Maguire CA, Kotin RM, Martin DR, Sena-Esteves M.Mol Ther. 2016 Aug;24(7):1247-57. doi: 10.1038/mt.2016.84. Epub 2016 Apr 27.
  2. The AAV9 Variant Capsid AAV-F Mediates Widespread Transgene Expression in Nonhuman Primate Spinal Cord After Intrathecal Administration. Beharry A, Gong Y, Kim JC, Hanlon KS, Nammour J, Hieber K, Eichler F, Cheng M, Stemmer-Rachamimov A, Stankovic KM, Welling DB, Ng C, Maguire CA.Hum Gene Ther. 2022 Jan;33(1-2):61-75. doi: 10.1089/hum.2021.069. Epub 2021 Aug 26.
  3. Engineered AAVs for non-invasive gene delivery to rodent and non-human primate nervous systems. Chen X, Ravindra Kumar S, Adams CD, Yang D, Wang T, Wolfe DA, Arokiaraj CM, Ngo V, Campos LJ, Griffiths JA, Ichiki T, Mazmanian SK, Osborne PB, Keast JR, Miller CT, Fox AS, Chiu IM, Gradinaru V.Neuron. 2022 Jul 20;110(14):2242-2257.e6. doi: 10.1016/j.neuron.2022.05.003. Epub 2022 May 27.
  4. Intravenous functional gene transfer throughout the brain of non-human primates using AAV. Chuapoco MR, Flytzanis NC, Goeden N, Octeau JC, Roxas KM, Chan KY, Scherrer J, Winchester J, Blackburn RJ, Campos LJ, Man KNM, Sun J, Chen X, Lefevre A, Singh VP, Arokiaraj CM, Shaya TF, Vendemiatti J, Jang MJ, Mich J, Bishaw Y, Gore B, Omstead V, Taskin N, Weed N, Ting J, Miller CT, Deverman BE, Pickel J, Tian L, Fox AS, Gradinaru V.Res Sq [Preprint]. 2023 Jan 13:rs.3.rs-1370972. doi: 10.21203/rs.3.rs-1370972/v1.
  5. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems Ken Y Chan, Min J Jang, Bryan B Yoo, Alon Greenbaum, Namita Ravi, Wei-Li Wu, Luis Sánchez-Guardado, Carlos Lois, Sarkis K Mazmanian, Benjamin E Deverman, Viviana Gradinaru. Published in final edited form as: Nat Neurosci. 2017 Aug; 20(8): 1172–1179.
  6. Multiplexed Cre-dependent selection yields systemic AAVs for targeting distinct brain cell types.Sripriya Ravindra Kumar, Timothy F. Miles, Xinhong Chen, David Brown, Tatyana Dobreva, Qin Huang, Xiaozhe Ding, Yicheng Luo, Pétur H. Einarsson, Alon Greenbaum, Min J. Jang, Benjamin E. Deverman, Viviana Gradinaru Nat Methods. Author manuscript; available in PMC 2020 Oct 20. Published in final edited form as: Nat Methods. 2020 May; 17(5): 541–550. Published online 2020 Apr 20. doi: 10.1038/s41592-020-0799-7
  7. AAV capsid variants with brain-wide transgene expression and decreased liver targeting after intravenous delivery in mouse and marmoset.Goertsen D, Flytzanis NC, Goeden N, Chuapoco MR, Cummins A, Chen Y, Fan Y, Zhang Q, Sharma J, Duan Y, Wang L, Feng G, Chen Y, Ip NY, Pickel J, Gradinaru V.Nat Neurosci. 2022 Jan;25(1):106-115. doi: 10.1038/s41593-021-00969-4. Epub 2021 Dec 9.PMID: 34887588
  8. Preclinical biodistribution, tropism, and efficacy of oligotropic AAV/Olig001 in a mouse model of congenital white matter disease.Francis JS, Markov V, Wojtas ID, Gray S, McCown T, Samulski RJ, Figueroa M, Leone P.Mol Ther Methods Clin Dev. 2021 Jan 21;20:520-534. doi: 10.1016/j.omtm.2021.01.009. eCollection 2021 Mar 12.
AAV Scale

1X10^12 VG/Capsid, 2X10^12 VG/Capsid, 5X10^12 VG/Capsid, 10X10^12 VG/Capsid

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