AAV Capsid Barcode Kit-Lung

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 sc-AAV-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-A signal. NGS technology is used to analyze the barcode data after AAV infection.  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

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

References:

1.Successful expansion but not complete restriction of tropism of adeno-associated virus by in vivo biopanning of random virus display peptide libraries.Michelfelder S, Kohlschütter J, Skorupa A, Pfennings S, Müller O, Kleinschmidt JA, Trepel M.PLoS One. 2009;4(4):e5122. doi: 10.1371/journal.pone.0005122. Epub 2009 Apr 9.

2.Pulmonary Targeting of Adeno-associated Viral Vectors by Next-generation Sequencing-guided Screening of Random Capsid Displayed Peptide Libraries. Körbelin J, Sieber T, Michelfelder S, Lunding L, Spies E, Hunger A, Alawi M, Rapti K, Indenbirken D, Müller OJ, Pasqualini R, Arap W, Kleinschmidt JA, Trepel M.Mol Ther. 2016 Jun;24(6):1050-1061. doi: 10.1038/mt.2016.62. Epub 2016 Mar 28.

5.A lung tropic AAV vector improves survival in a mouse model of surfactant B deficiency. Kang MH, van Lieshout LP, Xu L, Domm JM, Vadivel A, Renesme L, Mühlfeld C, Hurskainen M, Mižíková I, Pei Y, van Vloten JP, Thomas SP, Milazzo C, Cyr-Depauw C, Whitsett JA, Nogee LM, Wootton SK, Thébaud B.Nat Commun. 2020 Aug 6;11(1):3929. doi: 10.1038/s41467-020-17577-8.

6. Repeat Dosing of AAV2.5T to Ferret Lungs Elicits an Antibody Response That Diminishes Transduction in an Age-Dependent Manner.. Tang Y, Yan Z, Lin S, Huntemann ED, Feng Z, Park SY, Sun X, Yuen E, Engelhardt JF.Mol Ther Methods Clin Dev. 2020 Sep 16;19:186-200. doi: 10.1016/j.omtm.2020.09.008. eCollection 2020 Dec 11.

7. 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.

8.Genetic Modification of the AAV5 Capsid with Lysine Residues Results in a Lung-Tropic Liver-Detargeted Gene Transfer Vector. Stiles KM, Frenk EZ, Kaminsky SM, Crystal RG.Hum Gene Ther. 2022 Feb;33(3-4):148-154. doi: 10.1089/hum.2021.200.