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Precision Fluorescent Labeling of an Adeno‐Associated Virus Vector to Monitor the Viral Infection Pathway
Author(s) -
Zhang Chuanling,
Zhou Xueying,
Yao Tianzhuo,
Tian Zhenyu,
Zhou Demin
Publication year - 2018
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201700374
Subject(s) - bioorthogonal chemistry , transduction (biophysics) , intracellular , adeno associated virus , virus , capsid , gene delivery , computational biology , biology , green fluorescent protein , viral vector , virology , vector (molecular biology) , microbiology and biotechnology , click chemistry , gene , chemistry , transfection , biophysics , genetics , combinatorial chemistry , recombinant dna
Adeno‐associated virus 2 (AAV2) is a common vehicle for the delivery of a variety of therapeutic genes. A better understanding of the process of infection of AAV2 will advance our knowledge of AAV2 biology and allow for the optimization of AAV2 capsids with favorable transduction profiles. However, the precise fluorescent labeling of an AAV2 vector for probing virus tracking without affecting the nature of the virus remains a challenge. In this study, a lab‐synthesized azide‐moieties on the viral capsid at modifiable sites is precisely displayed. Upon bioorthogonal copper‐less click reaction, fluorophores are subsequently conjugated to AAV2 vectors for visualization of particles. Using this principle, the authors demonstrate that it can be used for visibly studying the cell entry, and intracellular trafficking of AAV2 particles, enabling the monitoring of the intracellular dynamics of AAV2 infection. This study provides new insights into the precision labeling of AAV2 particles with important implications for a better understanding of the molecular mechanism of therapeutic gene delivery.