Using In-situ Hybridization to Localize FAM171B Expression
Location
CSU Ballroom
Start Date
18-4-2016 10:00 AM
End Date
18-4-2016 11:30 AM
Student's Major
Biological Sciences
Student's College
Science, Engineering and Technology
Mentor's Name
Geoffrey Goellner
Mentor's Department
Biological Sciences
Mentor's College
Science, Engineering and Technology
Second Mentor's Name
Ashani Sudasinghe
Second Mentor's Department
Biological Sciences
Second Mentor's College
Science, Engineering and Technology
Description
The neurodegenerative disorder Huntington’s disease (HD) is a progressive and fatal disorder for which no cure exists (Orr & Zoghbi, 2000). The disease contains repeats of CAG (amino acid glutamine-Q) in their primary amino acid sequences (Ross, 2002). In afflicted individuals, these “polyglutamine” (polyQ) stretches expand beyond their normal ranges, leading to brain cell death (Orr & Zoghbi, 2000), and are thus inferred to be the main cause of pathological effects of HD (Ross, 2002). An uncharacterized protein called FAM171B also contains a polyQ stretch and is believed to perhaps function within the brain (Hypothetical Protein KIAA1946, 2010). The purpose of this research project is to gain understanding of where in the body FAM171B is expressed. If FAM171B is indeed found in the brain, it could be considered a candidate gene for a currently unidentified neurodegenerative disease. In-situ hybridization is the technique we utilized to carry out our research. For our purposes, this technique was performed on mouse brain tissue (IACUC: 15-02) in order to test for FAM171B expression. A “probe” was prepared by sub-cloning FAM171B into a vector containing promotors, and using these promoters to produce “in vitro transcribed mRNA” containing complementary nucleotide base-pairs to that of the FAM171B. Addition of DIG-labeled bases essentially labels the “probe,” allowing us to see the complementary mRNA strand of FAM171B in tested tissues by “lighting up” after probe application. Upon completion of our experiments, we expect to have high quality data concerning where in the brain FAM171B is expressed.
Using In-situ Hybridization to Localize FAM171B Expression
CSU Ballroom
The neurodegenerative disorder Huntington’s disease (HD) is a progressive and fatal disorder for which no cure exists (Orr & Zoghbi, 2000). The disease contains repeats of CAG (amino acid glutamine-Q) in their primary amino acid sequences (Ross, 2002). In afflicted individuals, these “polyglutamine” (polyQ) stretches expand beyond their normal ranges, leading to brain cell death (Orr & Zoghbi, 2000), and are thus inferred to be the main cause of pathological effects of HD (Ross, 2002). An uncharacterized protein called FAM171B also contains a polyQ stretch and is believed to perhaps function within the brain (Hypothetical Protein KIAA1946, 2010). The purpose of this research project is to gain understanding of where in the body FAM171B is expressed. If FAM171B is indeed found in the brain, it could be considered a candidate gene for a currently unidentified neurodegenerative disease. In-situ hybridization is the technique we utilized to carry out our research. For our purposes, this technique was performed on mouse brain tissue (IACUC: 15-02) in order to test for FAM171B expression. A “probe” was prepared by sub-cloning FAM171B into a vector containing promotors, and using these promoters to produce “in vitro transcribed mRNA” containing complementary nucleotide base-pairs to that of the FAM171B. Addition of DIG-labeled bases essentially labels the “probe,” allowing us to see the complementary mRNA strand of FAM171B in tested tissues by “lighting up” after probe application. Upon completion of our experiments, we expect to have high quality data concerning where in the brain FAM171B is expressed.
Recommended Citation
Jones, Brooke. "Using In-situ Hybridization to Localize FAM171B Expression." Undergraduate Research Symposium, Mankato, MN, April 18, 2016.
https://cornerstone.lib.mnsu.edu/urs/2016/poster-session-A/2
