title: Recovery of cell cycle delay following targeted gene repair by oligonucleotides.
abstract: We have previously shown that activation of the homologous recombinational repair pathway leads to a block of cell division in corrected cells, possibly through the activity of checkpoint proteins Chk1 and Chk2. In this study, we examine the long-term impact of this stalling on the growth of cells that have enabled gene repair events. Using a mutated eGFP gene as an episomal reporter, we show that corrected (eGFP-positive) cells contain only a few active replication templates 2 weeks after electroporation, yet do not display an apoptotic or senescent phenotype. By 6 weeks after electroporation, cells resume active replication with a cell cycle profile that is comparable to that of the non-corrected (eGFP-negative) population. These results indicate that the initial stalling is transient and eGFP-positive cells eventually resume a normal phenotypic growth pattern, allowing for passaging and expansion in vitro.
authors.affiliation: Department of Biological Sciences, University of Delaware, Delaware Biotechnology Institute, 15 Innovation Way, Newark, DE 19711, USA.
authors.forename: Luciana
authors.initials: L
authors.lastname: Ferrara
authors.forename: Julia U
authors.initials: JU
authors.lastname: Engstrom
authors.forename: Timothy
authors.initials: T
authors.lastname: Schwartz
authors.forename: Hetal
authors.initials: H
authors.lastname: Parekh-Olmedo
authors.forename: Eric B
authors.initials: EB
authors.lastname: Kmiec
mesh_terms: D001483:Base Sequence; D002453:Cell Cycle / Q000187:drug effects*; D004260:DNA Repair; D004261:DNA Replication; D009841:Oligonucleotides / Q000494:pharmacology*
chemical_list: D009841:Oligonucleotides