Recovery of cell cycle delay following targeted gene repair by oligonucleotides.

DNA Repair (Amst)

Department of Biological Sciences, University of Delaware, Delaware Biotechnology Institute, 15 Innovation Way, Newark, DE 19711, USA.

Published: October 2007

AI Article Synopsis

  • The study investigates how activation of the homologous recombinational repair pathway affects cell growth and division in corrected cells.
  • Corrected cells initially experience a temporary halt in replication, showing few active replication templates, but do not undergo cell death or senescence.
  • Over time, these cells regain normal growth rates similar to non-corrected cells, indicating that the initial stalling is not permanent and they can continue to multiply in the lab.

Article 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.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2083256PMC
http://dx.doi.org/10.1016/j.dnarep.2007.04.007DOI Listing

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