Tuesday 29 December 2015

Studying Gene Expression

Studying Gene Expression:
Knowing the transcriptional activity of a gene can give valuable insight to the function of the protein it encodes and to the role it plays in an organism. Gene activity in the same individual can vary from tissue to tissue, between different developmental stages, or even from morning to night time. Gene activity is influenced by the activity of other genes and the proteins they encode.  Gene expression can change in response to outside factors, such as the environment or exposure of the organism to chemical substances, competitors, or pathogens.
The classical approach to measuring the activity of a gene has been to isolate messenger RNA (mRNA), design nucleic acid molecules complementary to the gene of interest, and use those to estimate the amount of mRNA of the gene of interest present at a given time in the organism. Traditionally, this has been done for one gene at a time.
Using extremely small capillaries to apply short pieces of DNA, each uniquely representing one gene. Up to 25,000 genes can be represented on a single conventional 1.5 cm x 5 cm slide. Using these microscopic arrays of DNA spots, researchers can assess the relative amount of mRNA in a sample of all 25,000 represented genes (called the target spots) in the same time that it used to take to analyse the activity of a single gene.
Such technological advances have revolutionized the way molecular bioscience is done and have sped up the rate of new discoveries. However, they have also led to the rapid acquisition of huge amounts of data that require the use of biostatistics for analysis and validation of the collected data. In practice, gene activity is assessed, by labelling mRNA that was extracted from an organism, with fluorescent dyes. The labelled mRNA, known as the “probe” is applied to the glass slide and allowed to bind to its complementary spot on the array. This process is called hybridization. Subsequently, the unbound mRNA is washed off the slide. The slide is scanned and the amount of fluorescently labelled mRNA bound to each spot is proportional to the activity of the gene it represents.
In most cases, software analysis is then used to determine how much of a signal is due to biologically relevant processes and how much is due to technical “noise”. 



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