Back to Search
Start Over
The Gene Targeting Approach of Small Fragment Homologous Replacement (SFHR) Alters the Expression Patterns of DNA Repair and Cell Cycle Control Genes
- Source :
- Molecular Therapy. Nucleic Acids, Molecular Therapy: Nucleic Acids, Vol 5, Iss C (2016)
- Publication Year :
- 2016
- Publisher :
- Elsevier BV, 2016.
-
Abstract
- Cellular responses and molecular mechanisms activated by exogenous DNA that invades cells are only partially understood. This limits the practical use of gene targeting strategies. Small fragment homologous replacement (SFHR) uses a small exogenous wild-type DNA fragment to restore the endogenous wild-type sequence; unfortunately, this mechanism has a low frequency of correction. In this study, we used a mouse embryonic fibroblast cell line with a stably integrated mutated gene for enhanced green fluorescence protein. The restoration of a wild-type sequence can be detected by flow cytometry analysis. We quantitatively analyzed the expression of 84 DNA repair genes and 84 cell cycle control genes. Peculiar temporal gene expression patterns were observed for both pathways. Different DNA repair pathways, not only homologous recombination, as well as the three main cell cycle checkpoints appeared to mediate the cellular response. Eighteen genes were selected as highly significant target/effectors of SFHR. We identified a wide interconnection between SFHR, DNA repair, and cell cycle control. Our results increase the knowledge of the molecular mechanisms involved in cell invasion by exogenous DNA and SFHR. Specific molecular targets of both the cell cycle and DNA repair machineries were selected for manipulation to enhance the practical application of SFHR.
- Subjects :
- 0301 basic medicine
Cell cycle checkpoint
DNA repair
arrayed gene expression
cell cycle control
gene targeting
SFHR
Molecular Medicine
Drug Discovery3003 Pharmaceutical Science
Cell cycle
Biology
03 medical and health sciences
chemistry.chemical_compound
Drug Discovery
Gene
Small fragment homologous replacement
Gene targeting
Arrayed gene expression
lcsh:RM1-950
Molecular biology
Cell biology
lcsh:Therapeutics. Pharmacology
030104 developmental biology
Settore MED/03 - Genetica Medica
chemistry
Original Article
Exogenous DNA
Homologous recombination
DNA
Subjects
Details
- ISSN :
- 21622531
- Volume :
- 5
- Database :
- OpenAIRE
- Journal :
- Molecular Therapy - Nucleic Acids
- Accession number :
- edsair.doi.dedup.....8132d59fad3355cb4d404e7298ef5a4b