Oligonucleotide conjugated multi-functional adeno-associated viruses

Abstract Recombinant adeno-associated viruses (AAVs) are among the most commonly used vehicles for in vivo gene delivery. However, their tropism is limited, and additionally their efficacy can be negatively affected by prevalence of neutralizing antibodies in sera. Methodologies to systematically en...

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Autores principales:	Dhruva Katrekar, Ana M. Moreno, Genghao Chen, Atharv Worlikar, Prashant Mali
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3e63cac820274427a3c7a71f9e4eb6ba
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spelling	oai:doaj.org-article:3e63cac820274427a3c7a71f9e4eb6ba2021-12-02T15:09:07ZOligonucleotide conjugated multi-functional adeno-associated viruses10.1038/s41598-018-21742-x2045-2322https://doaj.org/article/3e63cac820274427a3c7a71f9e4eb6ba2018-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-21742-xhttps://doaj.org/toc/2045-2322Abstract Recombinant adeno-associated viruses (AAVs) are among the most commonly used vehicles for in vivo gene delivery. However, their tropism is limited, and additionally their efficacy can be negatively affected by prevalence of neutralizing antibodies in sera. Methodologies to systematically engineer AAV capsid properties would thus be of great relevance. In this regard, we develop here multi-functional AAVs by engineering precision tethering of oligonucleotides onto the AAV surface, and thereby enabling a spectrum of nucleic-acid programmable functionalities. Towards this, we engineered genetically encoded incorporation of unnatural amino acids (UAA) bearing bio-orthogonal chemical handles onto capsid proteins. Via these we enabled site-specific coupling of oligonucleotides onto the AAV capsid surface using facile click chemistry. The resulting oligo-AAVs could be sequence specifically labeled, and also patterned in 2D using DNA array substrates. Additionally, we utilized these oligo conjugations to engineer viral shielding by lipid-based cloaks that efficaciously protected the AAV particles from neutralizing serum. We confirmed these ‘cloaked AAVs’ retained full functionality via their ability to transduce a range of cell types, and also enable robust delivery of CRISPR-Cas9 effectors. Taken together, we anticipate this programmable oligo-AAV system will have broad utility in synthetic biology and AAV engineering applications.Dhruva KatrekarAna M. MorenoGenghao ChenAtharv WorlikarPrashant MaliNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-8 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Dhruva Katrekar Ana M. Moreno Genghao Chen Atharv Worlikar Prashant Mali Oligonucleotide conjugated multi-functional adeno-associated viruses
description	Abstract Recombinant adeno-associated viruses (AAVs) are among the most commonly used vehicles for in vivo gene delivery. However, their tropism is limited, and additionally their efficacy can be negatively affected by prevalence of neutralizing antibodies in sera. Methodologies to systematically engineer AAV capsid properties would thus be of great relevance. In this regard, we develop here multi-functional AAVs by engineering precision tethering of oligonucleotides onto the AAV surface, and thereby enabling a spectrum of nucleic-acid programmable functionalities. Towards this, we engineered genetically encoded incorporation of unnatural amino acids (UAA) bearing bio-orthogonal chemical handles onto capsid proteins. Via these we enabled site-specific coupling of oligonucleotides onto the AAV capsid surface using facile click chemistry. The resulting oligo-AAVs could be sequence specifically labeled, and also patterned in 2D using DNA array substrates. Additionally, we utilized these oligo conjugations to engineer viral shielding by lipid-based cloaks that efficaciously protected the AAV particles from neutralizing serum. We confirmed these ‘cloaked AAVs’ retained full functionality via their ability to transduce a range of cell types, and also enable robust delivery of CRISPR-Cas9 effectors. Taken together, we anticipate this programmable oligo-AAV system will have broad utility in synthetic biology and AAV engineering applications.
format	article
author	Dhruva Katrekar Ana M. Moreno Genghao Chen Atharv Worlikar Prashant Mali
author_facet	Dhruva Katrekar Ana M. Moreno Genghao Chen Atharv Worlikar Prashant Mali
author_sort	Dhruva Katrekar
title	Oligonucleotide conjugated multi-functional adeno-associated viruses
title_short	Oligonucleotide conjugated multi-functional adeno-associated viruses
title_full	Oligonucleotide conjugated multi-functional adeno-associated viruses
title_fullStr	Oligonucleotide conjugated multi-functional adeno-associated viruses
title_full_unstemmed	Oligonucleotide conjugated multi-functional adeno-associated viruses
title_sort	oligonucleotide conjugated multi-functional adeno-associated viruses
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/3e63cac820274427a3c7a71f9e4eb6ba
work_keys_str_mv	AT dhruvakatrekar oligonucleotideconjugatedmultifunctionaladenoassociatedviruses AT anammoreno oligonucleotideconjugatedmultifunctionaladenoassociatedviruses AT genghaochen oligonucleotideconjugatedmultifunctionaladenoassociatedviruses AT atharvworlikar oligonucleotideconjugatedmultifunctionaladenoassociatedviruses AT prashantmali oligonucleotideconjugatedmultifunctionaladenoassociatedviruses
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Oligonucleotide conjugated multi-functional adeno-associated viruses

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