Oligonucleotide conjugated multi-functional adeno-associated viruses

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 AA...

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Bibliographic Details
Published in:Scientific reports, Vol. 8, No. 1 (2018), p. 3589
Main Author: Katrekar, Dhruva
Other Involved Persons: Moreno, Ana M ; Chen, Genghao ; Worlikar, Atharv ; Mali, Prashant
Format: electronic Article
Language:English
ISSN:2045-2322
Item Description:Date Completed 21.10.2019
Date Revised 22.10.2019
published: Electronic
Citation Status MEDLINE
Copyright: From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Physical Description:Online-Ressource
DOI:10.1038/s41598-018-21742-x
Subjects:
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520 |a 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 
611 2 7 |a Journal Article  |2 gnd 
611 2 7 |a Research Support, N.I.H., Extramural  |2 gnd 
611 2 7 |a Research Support, Non-U.S. Gov't  |2 gnd 
653 2 |a Amino Acids  |6 D000596  |a genetics  |6 Q000235 
653 2 |a Animals  |6 D000818 
653 2 |a Antibodies, Neutralizing  |6 D057134  |a blood  |6 Q000097  |a immunology  |6 Q000276 
653 2 |a CRISPR-Associated Protein 9  |6 D000076987  |a chemistry  |6 Q000737 
653 2 |a Capsid Proteins  |6 D036022  |a chemistry  |6 Q000737 
653 2 |a Click Chemistry  |6 D057930  |a methods  |6 Q000379 
653 2 |a Dependovirus  |6 D000229  |a *chemistry  |6 Q000737 
653 2 |a Genetic Therapy  |6 D015316  |a methods  |6 Q000379 
653 2 |a Genetic Vectors  |6 D005822  |a *chemistry  |6 Q000737 
653 2 |a HEK293 Cells  |6 D057809 
653 2 |a Humans  |6 D006801 
653 2 |a Oligonucleotides  |6 D009841  |a *chemistry  |6 Q000737 
653 2 |a Swine  |6 D013552  |a blood  |6 Q000097 
653 2 |a Transduction, Genetic  |6 D014161  |a *methods  |6 Q000379 
653 2 |a Transfection  |6 D014162  |a methods  |6 Q000379 
655 7 |a Amino Acids  |2 gnd 
655 7 |a Antibodies, Neutralizing  |2 gnd 
655 7 |a Capsid Proteins  |2 gnd 
655 7 |a Oligonucleotides  |2 gnd 
655 7 |a CRISPR-Associated Protein 9  |2 gnd 
655 7 |a EC 3.1.-  |2 gnd 
689 0 0 |A f  |a Journal Article 
689 0 1 |A f  |a Research Support, N.I.H., Extramural 
689 0 2 |A f  |a Research Support, Non-U.S. Gov't 
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689 2 0 |a Amino Acids 
689 2 1 |a Antibodies, Neutralizing 
689 2 2 |a Capsid Proteins 
689 2 3 |a Oligonucleotides 
689 2 4 |a CRISPR-Associated Protein 9 
689 2 5 |A r  |a EC 3.1.- 
689 2 |5 DE-601 
700 1 |a Moreno, Ana M 
700 1 |a Chen, Genghao 
700 1 |a Worlikar, Atharv 
700 1 |a Mali, Prashant 
773 0 8 |i in  |t Scientific reports  |g Vol. 8, No. 1 (2018), p. 3589  |q 8:1<3589  |w (DE-601)NLM212555170  |x 2045-2322 
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