Abstract
Fluorescent-based visualization techniques have long been used to monitor biological activity. This chapter explores the delivery of reporter genes as a means to assay and track activity in biological systems. Bioluminescence is the production of light due to biochemical processes. By encoding genes for bioluminescence, biological processes can be visualized based on gene expression. This chapter also discusses the primary applications of bioluminescence as seen through bioluminescent imaging techniques, flow cytometry, and PCR-based methods of gene detection. These techniques are described in terms of researching gene expression, cancer therapy, and protein interactions.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Abbady AQ, Twair A, Ali B, Murad H (2017) Characterization of annexin V fusion with the superfolder GFP in liposomes binding and apoptosis detection. Front Physiol. https://doi.org/10.3389/fphys.2017.00317
Alauddin MM, Gelovani JG (2010) Radiolabeled nucleoside analogues for PET imaging of HSV1-tk gene expression. Curr Top Med Chem 10:1617–1632. https://doi.org/10.2174/156802610793176756
Alberts B, Johnson A, Lewis J et al (2002) Molecular biology of the cell, 4th edn. Garland Science, New York. https://doi.org/10.1091/mbc.E14-10-1437
Alwine JC, Kemp DJ, Stark GR (1977) Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A 74:5350–5354. https://doi.org/10.1073/pnas.74.12.5350
Andersen CL, Jensen JL, Ørntoft TF (2004) Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 64:5245–5250. https://doi.org/10.1158/0008-5472.CAN-04-0496
Bakayan A, Domingo B, Miyawaki A, Llopis J (2015) Imaging Ca2+ activity in mammalian cells and zebrafish with a novel red-emitting aequorin variant. Pflugers Arch - Eur J Physiol 467(9):2031–2042. https://doi.org/10.1007/s00424-014-1639-3
Benaron DA, Contag PR, Contag CH (1997) Imaging brain structure and function, infection and gene expression in the body using light. Philos Trans R Soc Lond Ser B Biol Sci 352(1354):755–761. https://doi.org/10.1098/rstb.1997.0059
Bhaumik S, Lewis XZ, Gambhir SS (2004) Optical imaging of Renilla luciferase, synthetic Renilla luciferase, and firefly luciferase reporter gene expression in living mice. J Biomed Opt 9(3):578–586. https://doi.org/10.1117/1.1647546
Bonner WA, Hulett HR, Sweet RG, Herzenberg LA (1972) Fluorescence activated cell sorting. Rev Sci Instrum 43:404–409. https://doi.org/10.1063/1.1685647
Boute N, Pernet K, Issad T (2001) Monitoring the activation state of the insulin receptor using bioluminescence resonance energy transfer. Mol Pharmacol 60(4):640–645. https://doi.org/10.1124/mol.106.026989.lin
Boute N, Jockers R, Issad T (2002) The use of resonance energy transfer in high-throughput screening: BRET versus FRET. Trends Pharmacol Sci 23:351–354. https://doi.org/10.1016/S0165-6147(02)02062-X
Brazma A, Hingamp P, Quackenbush J et al (2001) Minimum information about a microarray experiment (MIAME)—toward standards for microarray data. Nat Genet 29:365–371. https://doi.org/10.1038/ng1201-365
Brogan J, Li F, Li W et al (2012) Imaging molecular pathways: reporter genes. Radiat Res 177:508–513. https://doi.org/10.1667/RR2918.1
Bullok K, Piwnica-Worms D (2005) Synthesis and characterization of a small, membrane-permeant, caspase-activatable far-red fluorescent peptide for imaging apoptosis. J Med Chem 48:5404–5407. https://doi.org/10.1021/jm050008p
Bustin SA, Benes V, Nolan T, Pfaffl MW (2005) Quantitative real-time RT-PCR - a perspective. J Mol Endocrinol 34:597–601. https://doi.org/10.1677/jme.1.01755
Bustin SA, Benes V, Garson JA et al (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55:611–622. https://doi.org/10.1373/clinchem.2008.112797
Chalfie M, Tu Y, Euskirchen G et al (1994) Green fluorescent protein as a marker for gene expression. Sci 263:802–805. https://doi.org/10.1126/science.8303295
Chou CC, Huang YH (2012) Nucleic acid sandwich hybridization assay with quantum dot-induced fluorescence resonance energy transfer for pathogen detection. Sensors (Switzerland) 12:16660–16672. https://doi.org/10.3390/s121216660
Close DM, Xu T, Sayler GS, Ripp S (2011) In vivo bioluminescent imaging (BLI): noninvasive visualization and interrogation of biological processes in living animals. Sensors 11:180–206
Condeelis J, Weissleder R (2010) In vivo imaging in cancer. Cold Spring Harb Perspect Biol. https://doi.org/10.1101/cshperspect.a003848
Contag CH, Contag PR, Mullins JI, Spilman SD, Stevenson DK, Benaron DA (1995) Photonic detection of bacterial pathogens in living hosts. Mol Microbiol 18(4):593–603. https://doi.org/10.1111/j.1365-2958.1995.mmi_18040593.x
Contag CH, Ross BD (2002) It’s not just about anatomy: in vivo bioluminescence imaging as an eyepiece into biology. J Magn Reson Imag 16:378–387. https://doi.org/10.1002/jmri.10178
Cronin M, Akin AR, Collins SA, Meganck J, Kim JB, Baban CK et al (2012) High resolution in vivo bioluminescent imaging for the study of bacterial tumour targeting. PLoS One 7(1). https://doi.org/10.1371/journal.pone.0030940
Davey HM, Kell DB (1996) Flow cytometry and cell sorting of heterogeneous microbial populations: the importance of single-cell analyses. Microbiol Rev 60:641–696. https://doi.org/10.1016/j.mimet.2004.10.020
Deluca M, McElroy WD (1974) Kinetics of the firefly luciferase catalyzed reactions. Biochemistry 13(5):921–925. https://doi.org/10.1021/bi00702a015
Deluca M, McElroy WD (1978) [1] purification and properties of firefly luciferase. Methods Enzymol 57:3–15. https://doi.org/10.1016/0076-6879(78)57003-1
Deroose CM, Reumers V, Gijsbers R et al (2006) Noninvasive monitoring of long-term lentiviral vector-mediated gene expression in rodent brain with bioluminescence imaging. Mol Ther 14:423–431. https://doi.org/10.1016/j.ymthe.2006.05.007
Fraga H (2008) Firefly luminescence: a historical perspective and recent developments. Photochem Photobiol Sci 7:146–158. https://doi.org/10.1039/b719181b
Gilad AA, Ziv K, Mcmahon MT et al (2008) MRI reporter genes. J Nucl Med 49:1905–1908. https://doi.org/10.2967/jnumed.108.053520
Gilad AA, Van Laarhoven HWM, Mcmahon MT et al (2009) Feasibility of concurrent dual contrast enhancement using CEST contrast agents and superparamagnetic iron oxide particles. Magn Reson Med 61:970–974. https://doi.org/10.1002/mrm.21928
Givan A (2011) Flow cytometry: an introduction. Methods Mol Biol 699:1–29. https://doi.org/10.1007/978-1-61737-950-5_1
Gould SJ, Subramani S (1988) Firefly luciferase as a tool in molecular and cell biology. Anal Biochem 175:5–13. https://doi.org/10.1016/0003-2697(88)90353-3
Hasegawa S, Furukawa T, Saga T (2010) Molecular MR imaging of cancer gene therapy: ferritin transgene reporter takes the stage. Magn Reson Med Sci 9:37–47. https://doi.org/10.2463/mrms.9.37
Herzenberg LA, Sweet RG, Herzenberg LA (1976) Fluorescence-activated cell sorting. Sci Am 234:108–117. https://doi.org/10.1038/scientificamerican0376-108
Herzenberg LA, Tung J, Moore WA et al (2006) Interpreting flow cytometry data: a guide for the perplexed. Nat Immunol 7:681–685. https://doi.org/10.1038/ni0706-681
Higuchi-Sanabria R, Garcia EJ, Tomoiaga D et al (2016) Characterization of fluorescent proteins for three- and four-color live-cell imaging in S. cerevisiae. PLoS One. https://doi.org/10.1371/journal.pone.0146120
Himmelreich U, Dresselaers T (2009) Cell labeling and tracking for experimental models using magnetic resonance imaging. Methods 48:112–124. https://doi.org/10.1016/j.ymeth.2009.03.020
Hofmann A (2010) Spectroscopic techniques: I. Spectrophotometric techniques. In: Principles and techniques of biochemistry and molecular biology. Cambridge University Press, Cambridge, pp 477–521
Holland PM, Abramson RD, Watson R, Gelfand DH (1991) Detection of specific polymerase chain reaction product by utilizing the 5’----3’ exonuclease activity of Thermus aquaticus DNA polymerase. Proc Natl Acad Sci U S A 88:7276–7280
Honigman A, Zeira E, Ohana P, Abramovitz R, Tavor E, Bar I et al (2001) Imaging transgene expression in live animals. Mol Ther 4(3):239–249. https://doi.org/10.1006/mthe.2001.0437
Jacobs A, Tjuvajev JG, Dubrovin M, et al (2001) Positron emission tomography-based imaging of transgene expression mediated by replication-conditional, oncolytic herpes simplex virus type 1 mutant vectors in vivo. Cancer Res 61:2983–2995. PubMed PMID: 11306477
Jahan-Tigh RR, Ryan C, Obermoser G, Schwarzenberger K (2012) Flow cytometry. J Invest Dermatol 132:1–6. https://doi.org/10.1038/jid.2012.282
Kain SR, Adams M, Kondepudi A et al (1995) Green fluorescent protein as a reporter of gene expression and protein localization. Biotechniques 19:650–655
Kang JH, Chung J-K (2008) Molecular-genetic imaging based on reporter gene expression. J Nucl Med 49:164S–179S
Keith M, Farrell P, Behie I, Behie L (2000) Use of flow cytometry to rapidly optimize the transfection of animal cells. Biotechniques 28:14–54
Lakowicz JR (2006) Fluorophores. In: Principles of fluoescence spectroscopy, 3rd edn. Springer, Berlin, pp 63–69
Li X, Zhao X, Fang Y et al (1998) Generation of destabilized green fluorescent protein as a transcription reporter. J Biol Chem 273:34970–34975. https://doi.org/10.1074/jbc.273.52.34970
Liu G, Swierczewska M, Niu G et al (2011) Molecular imaging of cell-based cancer immunotherapy. Mol Biosyst 7:993
Liu Z, Lavis LD, Betzig E (2015) Imaging live-cell dynamics and structure at the single-molecule level. Mol Cell 58:644. https://doi.org/10.1016/j.molcel.2015.02.033
Lorenz WW, McCann RO, Longiaru M, Cormier MJ (1991) Isolation and expression of a cDNA encoding Renilla reniformis luciferase. Proc Natl Acad Sci U S A 88(10):4438–4442. https://doi.org/10.1073/pnas.88.10.4438
Massoud TF, Gambhir SS (2003) Molecular imaging in living subjects: seeing fundamental biological processes in a new light. Genes Dev 17:545–580. https://doi.org/10.1101/gad.1047403
Matsumoto Y, Jasanoff A (2013) Metalloprotein-based MRI probes. FEBS Lett 587:1021–1029. https://doi.org/10.1016/j.febslet.2013.01.044
McElroy WD, Hastings JW, Coulombre J, Sonnenfeld V (1953) The mechanism of action of pyrophosphate in firefly luminescence. Arch Biochem Biophys 46(2):399–416. https://doi.org/10.1016/0003-9861(53)90211-2
Mempel TR, Pittet MJ, Khazaie K et al (2006) Regulatory T cells reversibly suppress cytotoxic T cell function independent of effector differentiation. Immunity 25:129–141. https://doi.org/10.1016/j.immuni.2006.04.015
Mezzanotte L, van t’ Root M, Karatas H, et al (2017) In vivo molecular bioluminescence imaging: new tools and applications. Trends Biotechnol 35:640–652. https://doi.org/10.1016/j.tibtech.2017.03.012
Minn I, Bar-Shir A, Yarlagadda K et al (2015) Tumor-specific expression and detection of a CEST reporter gene. Magn Reson Med 74:544–549. https://doi.org/10.1002/mrm.25748
Morin JG, Hastings JW (1971) Energy transfer in a bioluminescent system. J Cell Physiol 77(3):313–318. https://doi.org/10.1002/jcp.1040770305
Nafissi N, Slavcev R (2012) Construction and characterization of an in-vivo linear covalently closed DNA vector production system. Microb Cell Fact 11:154. https://doi.org/10.1186/1475-2859-11-154
Nafissi N, Alqawlaq S, Lee EA et al (2014) DNA ministrings: highly safe and effective gene delivery vectors. Mol Ther Nucleic Acids 3:e165. https://doi.org/10.1038/mtna.2014.16
Nagai T, Ibata K, Park ES et al (2002) A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications. Nat Biotechnol 20:87–90. https://doi.org/10.1038/nbt0102-87
Nolan T, Hands RE, S a B (2006) Quantification of mRNA using real-time RT-PCR. Nat Protoc 1:1559–1582. https://doi.org/10.1038/nprot.2006.236
Overbergh L, Giulietti A, Valckx D, et al (2003) The use of real-time reverse transcriptase PCR for the quantification of cytokine gene expression. J Biomol Tech 14:33–43. PMC2279895
Penheiter AR, Russell SJ, Carlson SK (2012) The sodium iodide symporter (NIS) as an imaging reporter for gene, viral, and cell-based therapies. Curr Gene Ther 12:33–47. https://doi.org/10.2174/156652312799789235
Permyakov E (1993) Luminescent spectroscopy of proteins. CRC Press, London
Piatkevich KD, Verkhusha VV (2011) Guide to red fluorescent proteins and biosensors for flow cytometry. Methods Cell Biol. https://doi.org/10.1016/B978-0-12-374912-3.00017-1
Pichler BJ, Judenhofer MS, Pfannenberg C (2008) Multimodal imaging approaches: PET/CT and PET/MRI. Handb Exp Pharmacol 185:109–132. https://doi.org/10.1007/978-3-540-72718-7-6
Rice BW, Cable MD, Nelson MB (2001) In vivo imaging of light-emitting probes. J Biomed Opt 6(4):432–440. https://doi.org/10.1117/1.1413210
Richards MP, Poch SM (2002) Quantitative analysis of gene expression by reverse transcription polymerase chain reaction and capillary electrophoresis with laser-induced fluorescence detection. Mol Biotechnol 21:19–37. https://doi.org/10.1385/MB:21:1:019
Rizzuto R, Brini M, Pizzo P et al (1995) Chimeric green fluorescent protein as a tool for visualizing subcellular organelles in living cells. Curr Biol 5:635–642. https://doi.org/10.1016/S0960-9822(95)00128-X
Robert S, Lacroix R, Poncelet P et al (2012) High-sensitivity flow cytometry provides access to standardized measurement of small-size microparticles-brief report. Arterioscler Thromb Vasc Biol 32:1054–1058. https://doi.org/10.1161/ATVBAHA.111.244616
Rodriguez A, Nabi IR, Meighen E (1985) ATP turnover by the fatty acid reductase complex of Photobacterium phosphoreum. Can J Biochem Cell Biol 63(10):1106–1111
Sadikot RT, Wudel JL, Jansen DE, Debelak JP, Yull FE, Christonan JW et al (2002) Hepatic cryoablation-induced multisystem injury: bioluminescent detection of NF-κB activation in a transgenic mouse model. J Gastrointest Surg 6(2):264–270. https://doi.org/10.1016/S1091-255X(01)00064-6
Sadikot RT, Blackwell TS (2005) Bioluminescence imaging. Proc Am Thorac Soc 2:511–540. doi: https://doi.org/10.1513/pats.200507-067DS
Sato A, Klaunberg B, Tolwani R (2004) In vivo bioluminescence imaging. Comp Med 54:631–634
Schulze A, Downward J (2001) Navigating gene expression using microarrays—a technology review. Nat Cell Biol 3:E190–E195. https://doi.org/10.1038/35087138
Seliger HH, McElroy WD (1960) Spectral emission and quantum yield of firefly bioluminescence. Arch Biochem Biophys 88(1):136–141. https://doi.org/10.1016/0003-9861(60)90208-3
Serganova I, Mayer-Kukuck P, Huang R, Blasberg R (2008) Molecular imaging: reporter gene imaging. Handb Exp Pharmacol 185:167–223. https://doi.org/10.1007/978-3-540-77496-9-8
Shaner NC, Lin MZ, McKeown MR et al (2008) Improving the photostability of bright monomeric orange and red fluorescent proteins. Nat Methods 5:545–551. https://doi.org/10.1038/nmeth.1209
Shcherbakova DM, Verkhusha VV (2013) Near-infrared fluorescent proteins for multicolor in vivo imaging. Nat Methods 10:751–754. https://doi.org/10.1038/nmeth.2521
Shimomura O (2006) Bioluminescence, Chemical principles and methods, pp 1–29. https://doi.org/10.1142/9789812773647
Slonim DK, Yanai I (2009) Getting started in gene expression microarray analysis. PLoS Comput Biol 5:e1000543. https://doi.org/10.1371/journal.pcbi.1000543
Soboleski MR, Oaks J, Halford WP (2005) Green fluorescent protein is a quantitative reporter of gene expression in individual eukaryotic cells. Fed Am Soc Exp Biol 19:440–442. https://doi.org/10.1096/fj.04-3180fje
Stacer AC, Nyati S, Moudgil P, Iyengar R, Luker KE, Rehemtulla A, Luker GD (2013) NanoLuc reporter for dual luciferase imaging in living animals. Mol Imaging 12(7). https://doi.org/10.2310/7290.2013.00062
Tangney M, Francis KP (2012) In vivo optical imaging in gene & cell therapy. Curr Gene Ther 12:2–11. https://doi.org/10.2174/156652312799789299
T.E. Consortium (2011/2016) Standards, guidelines and best practices for RNA-Seq V1.0 (June 2011). https://genome.ucsc.edu/ENCODE/protocols/dataStandards/ENCODE_RNAseq_Standards_V1.0.pdf
van Roessel P, Brand AH (2002) Imaging into the future: visualizing gene expression and protein interactions with fluorescent proteins. Nat Cell Biol 4:E15–E20. https://doi.org/10.1038/ncb0102-e15
Vandsburger MH, Radoul M, Cohen B, Neeman M (2013) MRI reporter genes: applications for imaging of cell survival, proliferation, migration and differentiation. NMR Biomed 26:872–884. https://doi.org/10.1002/nbm.2869
Velculescu VE, Zhang L, Vogelstein B, Kinzler KW (1995) Serial analysis of gene expression. Science 270:484–487
Waerzeggers Y, Monfared P, Viel T et al (2009) Methods to monitor gene therapy with molecular imaging. Methods 48:146–160
Waerzeggers Y, Viel T, Schäfers S et al (2013) Positron emission tomography-based molecular imaging. Neuromethods 77:301–327. https://doi.org/10.1007/7657_2012_36
Wang Z, Gerstein M, Snyder M (2009) RNA-Seq: a revolutionary tool for transcriptomics. Nat Rev Genet 10:57–63. https://doi.org/10.1038/nrg2484
Wang Z, Wang F, Hida N et al (2015) Design of a functional cyclic HSV1-TK reporter and its application to PET imaging of apoptosis. Nat Protoc 10:807–821. https://doi.org/10.1038/nprot.2015.048
Weissleder R, Moore a MU et al (2000) In vivo magnetic resonance imaging of transgene expression. Nat Med 6:351–355. https://doi.org/10.1038/73219
Wetterwald A, van der Pluijm G, Que I, Sijmons B, Buijs J, Karperien M, Löwik CW, Gautschi E, Thalmann GN, Cecchini MG (2002) Optical imaging of cancer metastasis to bone marrow: a mouse model of minimal residual disease. Am J Pathol 160(3):1143–1153. https://doi.org/10.1016/S0002-9440(10)64934-6
Widder EA (2010) Bioluminescence in the ocean: origins of biological, chemical, and ecological diversity. Science (80–) 704:704–708. doi: https://doi.org/10.1126/science.1174269
Williams RS, Johnston SA, Riedy M et al (1991) Introduction of foreign genes into tissues of living mice by DNA-coated microprojectiles. Proc Natl Acad Sci U S A 88:2726–2730. https://doi.org/10.1073/PNAS.88.7.2726
Williams SJ, Schwer C, Krishnarao AS et al (1996) Quantitative competitive polymerase chain reaction: analysis of amplified products of the HIV-1 gag gene by capillary electrophoresis with laser-induced fluorescence detection. Anal Biochem 236:146–152. https://doi.org/10.1006/abio.1996.0143
Wong ML, Medrano JF (2005) Real-time PCR for mRNA quantitation. BioTechniques 39(1):75–85
Yaghoubi SS, Gambhir SS (2006) PET imaging of herpes simplex virus type 1 thymidine kinase (HSV1-tk) or mutant HSV1-sr39tk reporter gene expression in mice and humans using [18F]FHBG. Nat Protoc 1:3069–3075. https://doi.org/10.1038/nprot.2006.459
Yaghoubi SS, Campbell DO, Radu CG, Czernin J (2012) Positron emission tomography reporter genes and reporter probes: gene and cell therapy applications. Theranostics 2:374–391. https://doi.org/10.7150/thno.3677
Youn H, Hong KJ (2012) In vivo noninvasive small animal molecular imaging. Osong Public Health Res Perspect 3:48–59. https://doi.org/10.1016/j.phrp.2012.02.002
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer International Publishing AG, part of Springer Nature
About this chapter
Cite this chapter
Slavcev, R.A., Sum, C.H., St. Jean, J., Huh, H., Nafissi, N. (2018). Specific Systems for Evaluation. In: Slavcev, R., Wettig, S., Zeng, Z. (eds) Nanomedicine. Experientia Supplementum, vol 110. Springer, Cham. https://doi.org/10.1007/978-3-319-78259-1_4
Download citation
DOI: https://doi.org/10.1007/978-3-319-78259-1_4
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-319-78258-4
Online ISBN: 978-3-319-78259-1
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)