Ch-Ch-Ch-Changes! Measuring the Differential Expression of Small Noncoding Rna in Influenza A H1n1 Infected Epithelial Cells Using Q-Pcr
Date of Award
2012
Document Type
Thesis
Degree Name
Bachelors
Department
Natural Sciences
First Advisor
Walstrom, Katherine
Keywords
Influenza, PCR, Molecular Biology, sncRNA, mRNA, Swine Flu, Biochemistry, Micro RNA, Small Nucleolar RNA
Area of Concentration
Biochemistry
Abstract
Micro and small nucleolar RNA (miRNA and snoRNA) represent two classes of functional small-noncoding RNA (sncRNA) whose expression is closely regulated within the cell (Holley and Topkara, 2011). Viral infection has been shown to influence the expression levels of these sncRNA. One such virus, influenza A H1N1, poses a threat to the current and future public health (Peng et al, 2011). Prior to this project, deep sequencing of the small RNA (<200 nt) extracted from normal human bronchial epithelial (NHBE) cells that were infected with influenza A H1N1 (2009 strain) was performed. By measuring sncRNA expression in a similar set of influenza infected cells using quantitative polymerase chain reaction (qPCR), this project sought to confirm previous observations of differentially expressed sncRNA. The small RNA extracted from uninfected and infected NHBE cells was converted to cDNA and quantified via qPCR. Primers were designed to target highly expressed fragments of genes that were shown to be differentially expressed in the deep sequencing results. A comparison of fold-changes measured by each technique yielded a Pearsons' correlation coefficient of 0.12, suggesting inherent differences between techniques and experimental error. Despite these issues, this project identified several sncRNA that are differentially expressed in response to influenza infection.
Recommended Citation
Schuster, Andrew, "Ch-Ch-Ch-Changes! Measuring the Differential Expression of Small Noncoding Rna in Influenza A H1n1 Infected Epithelial Cells Using Q-Pcr" (2012). Theses & ETDs. 4676.
https://digitalcommons.ncf.edu/theses_etds/4676
Rights
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