Large-Scale RNA Interference Screening to Identify Transcriptional Regulators of a Tumor Suppressor Gene.
RNA interference (RNAi) is a powerful research tool that can be used to silence the expression of a specific gene. In the past several years, RNAi has provided the opportunity to identify factors and pathways involved in complex biological processes by performing unbiased loss-of-function screens on a genome-wide scale. Here we describe a genome-wide RNAi screening strategy to identify factors that regulates epigenetic silencing of a specific tumor suppressor gene, using RASSF1A as an example. The approach we describe is a general RNAi screening strategy that can be applied to identify other factors that drive and/or maintain epigenetic modifications on specific genes, including cancer-related genes.
Department of Pathology, Yale University School of Medicine, 310 Cedar Street, 208023, New Haven, CT, 06518, USA.
Matteo
M
Forloni
Department of Pathology, Yale University School of Medicine, 310 Cedar Street, 208023, New Haven, CT, 06518, USA.
Thuy
T
Ho
Department of Pathology, Yale University School of Medicine, 310 Cedar Street, 208023, New Haven, CT, 06518, USA.
Lisha
L
Sun
Department of Pathology, Yale University School of Medicine, 310 Cedar Street, 208023, New Haven, CT, 06518, USA. Narendra.Wajapeyee@yale.edu.
Narendra
N
Wajapeyee
D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D017930:Genes, Reporter; D016147:Genes, Tumor Suppressor; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D017422:Sequence Analysis, DNA; D014162:Transfection
D034741:RNA, Small Interfering
Epigenetic silencing; Positive-selection screen; RNA interference; shRNA; siRNA