Experimental strategies to manipulate the cellular levels of the multifunctional factor CTCF.
Cellular homeostasis is the result of an intricate and coordinated combinatorial of biochemical and molecular processes. Among them is the control of gene expression in the context of the chromatin structure which is central for cell survival. Interdependent action of transcription factors, cofactors, chromatin remodeling activities, and three-dimensional organization of the genome are responsible to reach exquisite levels of gene expression. Among such transcription factors there is a subset of highly specialized nuclear factors with features resembling master regulators with a large variety of functions. This is turning to be the case of the multifunctional nuclear factor CCCTC-binding protein (CTCF) which is involved in gene regulation, chromatin organization, and three-dimensional conformation of the genome inside the cell nucleus. Technically its study has turned to be challenging, in particular its posttranscriptional interference by small interference RNAs. Here we describe three main strategies to downregulate the overall abundance of CTCF in culture cell lines.
Departamento de Genética Molecular, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Apartado Postal 70-242, México, DF, 04510, México.
Edgar
E
González-Buendía
Rosario
R
Pérez-Molina
Erandi
E
Ayala-Ortega
Georgina
G
Guerrero
Félix
F
Recillas-Targa
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000076246:CCCTC-Binding Factor; D002460:Cell Line; D015536:Down-Regulation; D024881:Drug Resistance, Bacterial; D005434:Flow Cytometry; D055785:Gene Knockdown Techniques; D005821:Genetic Techniques; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D012097:Repressor Proteins / Q000172:deficiency* / Q000235:genetics*; D014161:Transduction, Genetic
D000900:Anti-Bacterial Agents; D000076246:CCCTC-Binding Factor; C000619697:CTCF protein, human; D008164:Luminescent Proteins; D034741:RNA, Small Interfering; D012097:Repressor Proteins