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. 2021 Apr 23;18:85. doi: 10.1186/s12985-021-01555-7

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© The Author(s) 2021

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — Methods for the identification of AAV capsids. The AAV capsid is the primary determinant of cell/tissue tropism. Efforts to identify novel capsids with enhanced target specificity and low immunogenicity are divided into two broad categories: native/fossil capsid isolation and capsid engineering (rational design or directed evolution). Native/fossil capsid isolation is achieved through live virus isolation from tissue culture, AAV-specific PCR or genome mining. AAV capsid engineering relies upon modification of key structural and genetic elements via rational design or directed evolution. Rational design exploits known aspects of AAV biology and structure, often focusing on surface-exposed regions of the AAV capsid. Directed evolution couples library-generated AAV capsid diversity and a selection scheme to identify variants with distinct properties. Strategies to generate AAV capsid libraries include error-prone PCR, DNA shuffling, and random peptide insertion. The libraries are then screened in cell culture systems, animal models, or a combination of both. Figure created with BioRender.com