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Strategies for Expressing Transgenes that Exceed the Packaging Capacity of AAVs

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Beth Kenkel

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BioRender Team

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Description: This template was adapted from the original submission. Edits were made to enhance scientific accuracy, optimal usability and/or to meet industry-leading design standards for science communication. Adeno-associated viral (AAV) vectors are popular gene therapy tools, but their genome packaging limit is ~4.7 kb. To exceed this size, transgenes are split between two AAVs and the two halves joined by use of: transplicing via a splice site donor and acceptor, homologous recombination (HR) mediated by homologous arms (HA); or a hybrid of these two approaches. CRISPR-mediate HR donor templates can also be split and are fully incorporated following sequential HR.

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This template was adapted from the original submission. Edits were made to enhance scientific accuracy, optimal usability and/or to meet industry-leading design standards for science communication. Adeno-associated viral (AAV) vectors are popular gene therapy tools, but their genome packaging limit is ~4.7 kb. To exceed this size, transgenes are split between two AAVs and the two halves joined by use of: transplicing via a splice site donor and acceptor, homologous recombination (HR) mediated by homologous arms (HA); or a hybrid of these two approaches. CRISPR-mediate HR donor templates can also be split and are fully incorporated following sequential HR.

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