title: Fluorogenic 'click-on' dendrimer reporter for rapid profiling of cell proliferation.
abstract: The application of small molecule fluorescent reporters to monitor biological systems is limited by their poor water solubility and background fluorescence of these reporters. Herein, we describe the synthesis and testing of a fluorogenic 'click' dendrimer reporter to monitor cellular processes. The reporter system consists of a polyamidoamine (PAMAM) dendrimer conjugated with 3-azido-7-hydroxy coumarin. After the copper(I)-catalyzed azide-alkyne cycloaddition reaction ('click' reaction) with alkyne-derivatized target molecules, the natively non-fluorescent construct has a strong enhancement in fluorescence. This fluorogenic dendrimer reporter can be used to efficiently monitor biological processes and the specificity afforded by the 'click' reaction greatly reduces background noise and enhances assay flexibility. We used this fluorogenic dendrimer reporter to monitor incorporation of 5-ethynyl-2'-deoxyuridine (EdU) into newly synthesized DNA, as a surrogate marker of cellular proliferation. We anticipate that this new class of fluorogenic reporter can be used to monitor a wide array of molecules and lends itself to high-throughput profiling of biological systems.
authors: Michigan Nanotechnology Institute for Medicine and Biological Sciences, 4608 Chemistry, 930 N. University Ave., University of Michigan, Ann Arbor, MI 48109, USA.
authors: Sascha N
authors: SN
authors: Goonewardena
authors: Pascale R
authors: PR
authors: Leroueil
authors: Colleen
authors: C
authors: Gemborys
authors: Pawan
authors: P
authors: Tahiliani
authors: Sarah
authors: S
authors: Emery
authors: James R
authors: JR
authors: Baker
authors: Hong
authors: H
authors: Zong
mesh_terms: D001386:Azides / Q000737:chemistry; D049109:Cell Proliferation; D057930:Click Chemistry; D003374:Coumarins / Q000737:chemistry; D050091:Dendrimers / Q000138:chemical synthesis / Q000737:chemistry*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D006801:Humans; D007624:KB Cells; D015394:Molecular Structure; D011073:Polyamines / Q000737:chemistry; D012995:Solubility
chemical_list: C507184:3-azido-7-hydroxycoumarin; D001386:Azides; D003374:Coumarins; D050091:Dendrimers; D005456:Fluorescent Dyes; C531249:Poly(amidoamine); D011073:Polyamines