Abstract
Recent advances have identified new genetic markers associated with the inheritance of celiac disease. These non-HLA target regions remain to be fully categorized. Investigation of associated SNPs indicates that the causal variants may alter specific gene expression. Thus, closer examination of potential causal variants found within regulatory regions could provide data relating to the mechanistic association. Molecular cloning is an established fundamental tool that enables investigators to examine the differential potential at a variant site. In conjunction with reporter gene assays, SNPs affecting gene expression can be uncovered and contribute to our understanding of the underlying pathogenic mechanisms. This chapter outlines the protocols necessary to clone risk variants and transfect these constructs into a T cell line for reporter assay analysis.
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Acknowledgements
The authors acknowledge Science Foundation Ireland (SFI) grant 09/IN.1/B2640 to R.M.M. We would also like to thank Dr. Richard Anney, Dr. Graham Kenny, Dr. Emma Quinn, and Dr. Anthony W. Ryan for their assistance and advice.
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Molloy, B., McManus, R. (2015). Cloning Gene Variants and Reporter Assays. In: Ryan, A. (eds) Celiac Disease. Methods in Molecular Biology, vol 1326. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2839-2_12
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DOI: https://doi.org/10.1007/978-1-4939-2839-2_12
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2838-5
Online ISBN: 978-1-4939-2839-2
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