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.
Tian
Jun
J
Bioengineering Laboratory, Department of Chemical and Biological Engineering, University at Buffalo, State University of New York, 908 Furnas Hall, Amherst, NY 14260-4200, USA.
Alimperti
Stella
S
Lei
Pedro
P
Andreadis
Stelios T
ST
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