The generation of a ZBTB16-inducible expression system in the ACHN adenocarcinoma cell line

Abstract

Abstract Objectives To establish an inducible system for controlled ZBTB16 protein expression in a renal adenocarcinoma cell line. Methods The Tet-On® 3G Inducible Expression System from Clontech was used to generate a tetracycline-regulated ZBTB16 expression system in the ACHN renal adenocarcinoma cell line. Results The Tet-On® 3G system successfully controlled the basal level of ZBTB16 with no detectable protein expression when doxycycline was absent from the culture medium. The addition of doxycycline induced ZBTB16 protein expression with as low as 20 ng/ml of doxycycline. Conclusion The firm regulation of transcription factor expression in cancer cell models is very important in investigating their biological roles. Many transcription factors, such as ZBTB16, will carry on their biological activities even with minimal expression levels. The Tet-On® 3G system was essential in controlling the basal expression to prevent transcription leakage and providing a tightly regulated method for expressing ZBTB16 in a manner resembling physiological conditions.