Design and development of high bioluminescent resonance energy transfer efficiency hybrid-imaging constructs

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• 作者：Manoj Kumar^a ; ^b ; Letí ; cia Kovalski^a ; David Broyles^a ; Eric A. Hunt^a ; Pirouz Daftarian^a ; Emre Dikici^a ; Sylvia Daunert^a ; Sapna K. Deo^a ; ^{sdeo@med.miami.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Bioluminescence ; Bioluminescence resonance energy transfer ; Quantum dots ; Renilla luciferase ; Imaging construct
• 刊名：Analytical Biochemistry
• 出版年：2016
• 出版时间：1 April 2016
• 年：2016
• 卷：498
• 期：Complete
• 页码：1-7
• 全文大小：1577 K

文摘

Here we describe the design and construction of an imaging construct with high bioluminescent resonance energy transfer (BRET) efficiency that is composed of multiple quantum dots (QDs; λ_em = 655 nm) self-assembled onto a bioluminescent protein, Renilla luciferase (Rluc). This is facilitated by the streptavidin–biotin interaction, allowing the facile formation of a hybrid-imaging construct (HIC) comprising up to six QDs (acceptor) grafted onto a light-emitting Rluc (donor) core. The resulting assembly of multiple acceptors surrounding a donor permits this construct to exhibit high resonance energy transfer efficiency (∼64.8%). The HIC was characterized using fluorescence excitation anisotropy measurements and high-resolution transmission electron microscopy. To demonstrate the application of our construct, a generation-5 (G5) polyamidoamine dendrimer (PAMAM) nanocarrier was loaded with our HIC for in vitro and in vivo imaging. We envision that this design of multiple acceptors and bioluminescent donor will lead to the development of new BRET-based systems useful in sensing, imaging, and other bioanalytical applications.