25 years of PCR – from idea to subcellular expression profiling

Mikael Kubista

PCR was born from a great idea conceived by Kary Mullis and later refined into qPCR by Russ Higushi. Specificity was added by Ken Livak. The technique found immediate use in genetic engineering leading to several breakthrough innovations, and later PCR became the preferred platform for clinical diagnostics. The quantitative aspect of qPCR became particularly valuable in biological and medical research, and is today key technology in systems biology. Moreover, the extreme sensitivity of qPCR, allowing the detection of single molecular copies, and the relative ease of use, which allows qPCR to be integrated in a streamlined high throughput workflow, has led to the exciting area of single cell expression profiling. An astonishing heterogeneity in transcript levels among seemingly like cells has been found, while correlation between genes’ expression is a fingerprint of the type of cell. Most recently, qPCR tomography has been developed to measure intracellular mRNA profiles, revealing gradients of transcripts within the cell, preparing it for asymmetric cell division.
Anders Stahlberg, Daniel Andersson, Johan Aurelius, Maryam Faiz, Marcela Pekna, Mikael Kubista, Milos Pekny. Defining cell populations with single-cell gene expression profiling: correlations and identification of astrocyte subpopulations. Nucl. Acids Res. 1–12. doi:10.1093/nar/gkq1182 (2010).
Radek Sindelka, Monika Sidova, David Svec, Mikael Kubista. Spatial expression profiles in the Xenopus laevis oocytes measured with qPCR tomography. Methods 51:87-91(2010).
M. Bengtsson, A. Ståhlberg, P. Rorsman, and M. Kubista Gene expression profiling in single cells from the pancreatic islets of Langerhans reveals lognormal distribution of mRNA levels. Genome Research 15, 1388-1392 (2005).
Research Highlights in Nature Review Genetics 6, 1758 (2005).

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