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Titolo:
Noninvasive optical imaging of firefly luciferase reporter gene expressionin skeletal muscles of living mice
Autore:
Wu, JC; Sundaresan, G; Iyer, M; Gambhir, SS;
Indirizzi:
Univ Calif Los Angeles, Sch Med, Crump Inst Mol Imaging, Los Angeles, CA 90095 USA Univ Calif Los Angeles Los Angeles CA USA 90095 Los Angeles, CA 90095 USA Univ Calif Los Angeles, Sch Med, Dept Mol & Med Pharmacol, Los Angeles, CA90095 USA Univ Calif Los Angeles Los Angeles CA USA 90095 Los Angeles, CA90095 USA Univ Calif Los Angeles, Sch Med, Dept Med, Div Cardiol, Los Angeles, CA 90095 USA Univ Calif Los Angeles Los Angeles CA USA 90095 Los Angeles, CA 90095 USA Univ Calif Los Angeles, Sch Med, DOE Lab Struct Biol & Mol Med, Los Angeles, CA 90095 USA Univ Calif Los Angeles Los Angeles CA USA 90095 Los Angeles, CA 90095 USA Univ Calif Los Angeles, Sch Med, Dept Biomath, Los Angeles, CA 90095 USA Univ Calif Los Angeles Los Angeles CA USA 90095 Los Angeles, CA 90095 USA Univ Calif Los Angeles, Sch Med, Jonsson Comprehens Canc Ctr, Los Angeles,CA 90095 USA Univ Calif Los Angeles Los Angeles CA USA 90095 Los Angeles,CA 90095 USA
Titolo Testata:
MOLECULAR THERAPY
fascicolo: 4, volume: 4, anno: 2001,
pagine: 297 - 306
SICI:
1525-0016(200110)4:4<297:NOIOFL>2.0.ZU;2-8
Fonte:
ISI
Lingua:
ENG
Soggetto:
POSITRON-EMISSION-TOMOGRAPHY; HUMORAL IMMUNE-RESPONSES; IN-VIVO; ADENOVIRAL VECTORS; BIOLUMINESCENT REPORTER; SYSTEMIC DELIVERY; PLASMID DNA; THERAPY; ANIMALS; MOUSE;
Keywords:
adenovirus; skeletal muscle; luciferase; CCD camera; reporter gene imaging; gene therapy;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
46
Recensione:
Indirizzi per estratti:
Indirizzo: Gambhir, SS Univ Calif Los Angeles, Sch Med, Crump Inst Mol Imaging, Los Angeles, CA 90095 USA Univ Calif Los Angeles Los Angeles CA USA 90095 CA 90095 USA
Citazione:
J.C. Wu et al., "Noninvasive optical imaging of firefly luciferase reporter gene expressionin skeletal muscles of living mice", MOL THER, 4(4), 2001, pp. 297-306

Abstract

The ability to monitor reporter gene expression noninvasively offers significant advantages over current techniques such as postmortem tissue staining or enzyme activity assays. Here we demonstrate a novel method of repetitively tracking in vivo gene expression of firefly luciferase (FL) in skeletal muscles of mice using a cooled charged coupled device (CCD) camera. We first show that the cooled CCD camera provides consistent and reproducible results within +/- 8% standard deviation from mean values, and a detection sensitivity (range tested: 1 X 10(4) - 1 X 10(9) plaque forming units (pfu)) of 1 X 10(6) pfu of E1 -deleted adenovirus expressing FL driven by a cytomegalovirus promoter (Ad-CMV-PL). The duration and magnitude of adenoviral mediated (1 X 10(9) pfu) FL gene expression were then followed over time. FL gene expression in immunocompetent Swiss Webster mice peaks within the first 48 hours, falls by 98% after 20 days, and persists for > 150 days. In contrast, FL activity in nude mice remains elevated for > 110 days. Finally, transduced Swiss Webster and nude mice were sacrificed to show that the in vivo CCD signals correlate well with in vitro luciferase enzyme assays (r(2)= 0.91 and 0.96, respectively). Our findings demonstrate the ability of the cooled CCD camera to sensitively and noninvasively track the location, magnitude, and persistence of FL gene expression. Monitoring of gene therapy studies in small animals may be aided considerably with further extensions of this technique.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 25/01/20 alle ore 03:14:56