Identification of G1‐Regulated Genes in Normally Cycling Human Cells

Abstract

Obtaining synchronous cell populations is essential for studies of the cell cycle. Methods such as serum withdrawal or use of drugs which block cells at specific points in the cell cycle almost certainly alter cellular events upon re‐entry into the cell cycle. Here we use a robotic mitotic shake‐off apparatus to select cells in late telophase of mitosis. Using two separate microarray experiments we examined gene activity in early G1: one isolated RNA hourly and the second isolated RNA every 15 minutes. We have also included data obtained by a variety of experimental methods to verify synchrony of these cell populations, including BrdU uptake, FACS, and microarray analyses of histone gene activity as well as activity of stress response genes. Our approach enabled us to identify a unique set of genes whose functions are currently unknown, in cells progressing normally through the cell division cycle. This group of genes may contain future targets for drug development and treatment of human disease.