What is a microarray?
A microarray is a way that scientists are able to view gene expression of a particular fragment of DNA, but be able to do it on a 1-million fold basis. This is possible through cutting a cell, isolating the DNA contents, identifying all of your genes of interest and then generating a list of those genes which can then be used to create a screen of fluorescing proteins which is recorded by a computer, which measures a heat map and then displays large sections of bright or dark patches are displayed and overlaid like in the picture below. The bright green and red portions of the image show high expression and the dark green and black shows low gene expression in that particular cellular component.
Discussion
Though there is a large amount of information to be found using microarrays, nothing has currently been done with colorectal cancer. 14-3-3 sigma is part of a superfamily of proteins but it has the most direct link to cancer. However, since it has a huge role in tumor suppression, it has a genetic basis in 14 other cancers besides colorectal cancer, including breast and ovarian cancers, which have much more funding and research being done than colon based cancers.
Though it has been used in microarrays pertaining to other types of cancer based proteins and 14-3-3sigma in varied types of experiments, nothing was found on these datasets to be useful to my project as a whole. A protocol of this type could be created involving a microarray to study the expression of 14-3-3sigma. However since the mutation of 14-3-3sigma occurs in the promoter, the protein is not functional and will just degrade within the cell. There is no gradient of expression when the protein is mutated in this way.
The probable best way to determine what the gene expression of 14-3-3sigma would be to create a CRISPR-Cas9 protocol to determine where it is working in the cell and how it is functioning in different GO Terms, which is described in my specific aims here.
Though it has been used in microarrays pertaining to other types of cancer based proteins and 14-3-3sigma in varied types of experiments, nothing was found on these datasets to be useful to my project as a whole. A protocol of this type could be created involving a microarray to study the expression of 14-3-3sigma. However since the mutation of 14-3-3sigma occurs in the promoter, the protein is not functional and will just degrade within the cell. There is no gradient of expression when the protein is mutated in this way.
The probable best way to determine what the gene expression of 14-3-3sigma would be to create a CRISPR-Cas9 protocol to determine where it is working in the cell and how it is functioning in different GO Terms, which is described in my specific aims here.
References:
[1] DNA Microarray. (n.d.). Retrieved March 26, 2015, from http://learn.genetics.utah.edu/content/labs/microarray/
[1] DNA Microarray. (n.d.). Retrieved March 26, 2015, from http://learn.genetics.utah.edu/content/labs/microarray/