Candidate Gene Knockdown in Celiac Disease.
RNA interference (RNAi) is a powerful genetic tool that has created new opportunities in cell biology by allowing the specific modulation of gene expression under controlled conditions. Knockdown of genes associated with disease can provide valuable information pertaining to their function and potentially their role in the disease etiology. In the context of celiac disease, it allows us to examine closely the cellular changes that occur when the expression levels of genes of interest are reduced. Utilizing informative assays that demonstrate changes in cell behavior or other measurable endpoints such as cytokine production or migratory phenotypes can further our understanding of the pathogenic mechanisms in this prevalent autoimmune disorder. This chapter outlines protocols for examining the effects of RNAi on candidate genes and subsequent changes to migratory phenotype, transmigration, and adhesion.
Department of Medicine, Institute of Molecular Medicine, Trinity College Dublin, St. James's Hospital, Dublin 8, Ireland.
Ben
B
Molloy
Department of Medicine, Institute of Molecular Medicine, Trinity College Dublin, St. James's Hospital, Dublin 8, Ireland.
Michael
M
Freeley
Department of Medicine, Institute of Molecular Medicine, Trinity College Dublin, St. James's Hospital, Dublin 8, Ireland.
Aideen
A
Long
Department of Medicine, Institute of Molecular Medicine, Trinity College Dublin, St. James's Hospital, Dublin 8, Ireland. rmcmanus@tcd.ie.
Ross
R
McManus
D002446:Celiac Disease / Q000235:genetics*; D055785:Gene Knockdown Techniques; D006801:Humans; D034622:RNA Interference
Adhesion; Celiac disease; Migration; T cells; siRNA knockdown