Nascent RNA sequencing of unperturbed newly divided cells

Abstract: Establishing a definitive cell cycle progression has been one of the fundamental aims of cellular biology. Its importance lies in gaining insight into the basic processes of life as well as the functions of mutant cell cycle pathways in promoting cancer by replication deficiencies and loss of checkpoint control. Currently used methods to control cell cycle and synchronize cells, function by halting cell cycle progression. Such harsh methods are detrimental to the cell and insufficient to provide an accurate reflection of the cell cycle. This study focused on replicating and confirming the efficiency of a technique developed by Helmstetter, called the “Baby Machine,” that can produce new born cells with little to no perturbations. Using this in conjunction with a short pulse RNA labelling technique, called Bru-seq, allowed the capture and RNA sequencing of synchronized cells and its nascent RNA. Here we show the first glimpse into the transcriptional profile of newly divided cells as well as novel rapid exon splicing and transcription read-through processes.