Gaussia Luciferase Reporter Assay for Assessment of Gene Delivery Systems in Vivo

doi:10.1016/S1001-9294(10)60029-6

Abstract

Abstract: Objective To develop an alternative method for assessment of gene delivery systems in vivo. Methods Mouse primary spleen lymphocytes were genetically modified in vitro by a retroviral vector harboring a Gaussia luciferase (Gluc) expression cassette. After implantation of these cells into recipient mice, the expression of Gluc was detected in whole blood or plasma collected. Results As little as 10 uL whole blood drawn from the recipient mice could guarantee prompt reading of Gluc activity with a luminometer. And the reading was found in good correlation with the number of genetically modified spleen lymphocytes implanted to the mice. Conclusions Gluc may be useful as an in vivo reporter for gene therapy researches, and Gluc blood assay could provide an alternative method for assessment of gene delivery systems in vivo.

Key words: gene delivery systems, reporter, Gaussia luciferase

Funding:

Supported by National High Technology Research and De- velopment Program of China (863 Program) (2007AA021206, 2007AA021106)

Feng Chen, Zhen Xu, Jing Lu, Xiang Lü, Wen-li Mu, Ya-jun Wang, De-pei Liu and Chih-chuan Liang. Gaussia Luciferase Reporter Assay for Assessment of Gene Delivery Systems in Vivo[J].Chinese Medical Sciences Journal, 2010, 25(2): 95-99.

References 19

1.	Chalfie M, Tu Y, Euskirchen G, et al. Green fluorescent protein as a marker for gene expression. Science 1994; 263:802-5.
2.	Heim R, Cubitt AB, Tsien RY. Improved green fluorescence. Nature 1995; 373:663-4.
3.	Zolotukhin S, Potter M, Hauswirth WW, et al. A "humanized" green fluorescent protein cDNA adapted for high- level expression in mammalian cells. J Virol 1996; 70: 4646-54.
4.	Larochelle A, Choi U, Shou Y, et al. In vivo selection of hematopoietic progenitor cells and temozolomide dose intensification in rhesus macaques through lentiviral transduction with a drug resistance gene. J Clin Invest 2009; 119:1952-63.
5.	Chin JY, Kuan JY, Lonkar PS, et al. Correction of a splice-site mutation in the beta-globin gene stimulated by triplex-forming peptide nucleic acids. Proc Natl Acad Sci U S A 2008; 105:13514-9.
6.	Bao R, Connolly DC, Murphy M, et al. Activation of cancer-specific gene expression by the survivin promoter. J Natl Cancer Inst 2002; 94:522-8.
7.	Bao R, Selvakumaran M, Hamilton TC. Use of a surrogate marker (human secreted alkaline phosphatase) to monitor in vivo tumor growth and anticancer drug efficacy in ovarian cancer xenografts. Gynecol Oncol 2000; 78: 373-9.
8.	Hiramatsu N, Kasai A, Hayakawa K, et al. Secreted protein-based reporter systems for monitoring inflammatory events: critical interference by endoplasmic reticulum stress. J Immunol Methods 2006; 315:202-7.
9.	Meng Y, Kasai A, Hiramatsu N, et al. Real-time monitoring of mesangial cell-macrophage cross-talk using SEAP in vitro and ex vivo. Kidney Int 2005; 68:886-93.
10	Murata T, Yasukawa T, Shiku H, et al. Electrochemical single-cell gene-expression assay combining dielectrophoretic manipulation with secreted alkaline phosphatase reporter system. Biosens Bioelectron 2009; 25:913-9.
11	Nishijo K, Hosoyama T, Bjornson CR, et al. Biomarker system for studying muscle, stem cells, and cancer in vivo. FASEB J 2009; 23:2681-90.
12	Hiramatsu N, Kasai A, Meng Y, et al. Alkaline phosphatase vs luciferase as secreted reporter molecules in vivo. Anal Biochem 2005; 339:249-56.
13	Badr CE, Hewett JW, Breakefield XO, et al. A highly sensitive assay for monitoring the secretory pathway and ER stress. PLoS One 2007; 2:e571.
14	Shiraiwa T, Kaneto H, Miyatsuka T, et al. Establishment of a non-invasive mouse reporter model for monitoring in vivo pdx-1 promoter activity. Biochem Biophys Res Com- mun 2007; 361:739-44.
15	Tannous BA, Kim D, Fernandez JL, et al. Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo. Mol Ther 2005; 11:435-43.
16	Wurdinger T, Badr C, Pike L, et al. A secreted luciferase for ex vivo monitoring of in vivo processes. Nat Methods 2008; 5:171-3.
17	Hagani AB, Rivire I, Tan C, et al. Activation conditions determine susceptibility of murine primary T-lymphocytes to retroviral infection. J Gene Med 1999; 1:341-51.
18	Annenkov AE, Daly GM, Chernajovsky Y. Highly efficient gene transfer into antigen-specific primary mouse lymphocytes with replication-deficient retrovirus expressing the 10A1 envelope protein. J Gene Med 2002; 4:133-40.
19	Colin M, Moritz S, Schneider H, et al. Haemoglobin interferes with the ex vivo luciferase luminescence assay: consequence for detection of luciferase reporter gene expression in vivo. Gene Ther 2000; 7:1333-6.

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