Lentiviral microarrays for real-time monitoring of gene expression dynamics.
We developed scalable live-cell microarrays to measure gene expression dynamics in real time and in a high-throughput manner. To this end, we generated dual-promoter lentiviral vectors harboring a transcriptional regulatory element encoding for green fluorescence protein to monitor cell activation in response to exogenous stimuli and a constitutive promoter driving red fluorescence protein for internal signal normalization. Lentivirus preparations were immobilized in a microarray format and after transduction on the array surface target cells were treated with cytokines and interrogated in real time using automated fluorescence microscopy, providing rich dynamic information over a period of several days. Data normalization by red fluorescence intensity eliminated errors due to spot-to-spot variability in transduction efficiency or changes in cell proliferation upon cytokine treatment. These results suggest that the lentivirus microarray can monitor gene expression in real-time and high-throughput manner thereby providing a useful tool for quantitative measurements of gene expression dynamics.
Bioengineering Laboratory, Department of Chemical and Biological Engineering, University at Buffalo, State University of New York, 908 Furnas Hall, Amherst, NY 14260-4200, USA.
Jun
J
Tian
Stella
S
Alimperti
Pedro
P
Lei
Stelios T
ST
Andreadis
D016207:Cytokines / Q000494:pharmacology; D020869:Gene Expression Profiling / Q000295:instrumentation / Q000379:methods; D049452:Green Fluorescent Proteins / Q000096:biosynthesis*; D006367:HeLa Cells; D006801:Humans; D056656:Lab-On-A-Chip Devices; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D011401:Promoter Regions, Genetic
D016207:Cytokines; D049452:Green Fluorescent Proteins