Detecting changes in the relative expression of KRAS2 splice variants using polymerase colonies
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Published on: December 9, 2016
James A Butz1, Kim G Roberts, Jeremy S Edwards
1Chemical Engineering, University of Delaware, Newark, Delaware 19716, USA.
Abstract:
Because splice variants of a gene with multiple isoforms give rise to proteins with different functions, it seems plausible that changes in the expression levels of the splice variants could be a contributing factor to disease. In fact, recent examples in the literature clearly illustrate that altered expression levels of splice variants may play an important role in disease. Furthermore, these works demonstrate that changes in expression levels could potentially be used to (1) monitor disease progression, (2) diagnose disease, and/or (3) determine disease state. In this work an immobilized form of PCR, known as polony technology, was adapted to quantify the relative expression levels of splice variants. Specifically, the relative expression levels of the two splice variants of the oncogene K-ras, namely, K-RAS2A and K-RAS2B, were determined using polony technology.
Insights
Changes in gene splice variants can indicate disease. This study used polony technology to quantify splice variant expression levels for K-RAS2A and K-RAS2B, offering potential disease monitoring tools.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- Alternative splicing generates diverse protein isoforms from a single gene, impacting cellular functions.
- Altered expression of splice variants is increasingly recognized as a factor in disease pathogenesis.
- Quantifying splice variant levels is crucial for understanding their role in disease.
Purpose of the Study:
- To adapt polony technology for precise quantification of splice variant expression levels.
- To determine the relative expression levels of the K-RAS2A and K-RAS2B splice variants of the K-ras oncogene.
Main Methods:
- Development and application of immobilized PCR (polony technology).
- Quantification of relative expression levels of specific gene splice variants.
Main Results:
- Polony technology was successfully adapted to quantify splice variant expression.
- The relative expression levels of K-RAS2A and K-RAS2B were determined using this method.
Conclusions:
- Quantifying splice variant expression levels is feasible and valuable.
- Altered splice variant expression, such as for K-ras, may serve as biomarkers for disease monitoring, diagnosis, or state determination.
