Gene-array analysis of glioma cells after treatment with an anti-PKCalpha siRNA: a general protocol.
Post-transcriptional gene silencing is a powerful tool to reveal gene function. In this chapter, we have used gene-array technology to analyze changes in gene expression after specific targeting of the protein kinase Calpha isoform with small interfering RNAs (siRNAs). Expression profiles were investigated on a human gene array of 588 key genes involved in DNA synthesis, apoptosis, cell-cell communication, intracellular signal-transduction pathways, cell-surface molecules and transcription factors. The data indicate that siRNAs are an ideal tool for target validation, and show that RNA-dependent RNA polymerase is not required for RNAi in mammalian cells.
Leirdal
Marianne
M
Department of Immunology, Molecular Medicine Group, The Norwegian Radium Hospital, Oslo.
Sioud
Mouldy
M
D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D003850:Deoxyribonuclease I; D020868:Gene Silencing; D005910:Glioma / Q000188:drug therapy / Q000235:genetics*; D006801:Humans; D007202:Indicators and Reagents; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000235:genetics*; D051571:Protein Kinase C-alpha; D034741:RNA, Small Interfering / Q000627:therapeutic use*; D014162:Transfection / Q000379:methods
D018076:DNA, Complementary; D007202:Indicators and Reagents; D034741:RNA, Small Interfering; C496628:PRKCA protein, human; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D003850:Deoxyribonuclease I