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Establishment of a Rabbit Oct4 Promoter-Based EGFP Reporter System
- Source :
- PLoS ONE, Vol 9, Iss 10, p e109728 (2014), PLoS ONE
- Publication Year :
- 2014
- Publisher :
- Public Library of Science (PLoS), 2014.
-
Abstract
- Rabbits are commonly used as laboratory animal models to investigate human diseases and phylogenetic development. However, pluripotent stem cells that contribute to germline transmission have yet to be established in rabbits. The transcription factor Oct4, also known as Pou5f1, is considered essential for the maintenance of the pluripotency of stem cells. Hence, pluripotent cells can be identified by monitoring Oct4 expression using a well-established Oct4 promoter-based reporter system. This study developed a rabbit Oct4 promoter-based enhanced green fluorescent protein (EGFP) reporter system by transfecting pROP2-EGFP into rabbit fetal fibroblasts (RFFs). The transgenic RFFs were used as donor cells for somatic cell nuclear transfer (SCNT). The EGFP expression was detected in the blastocysts and genital ridges of SCNT fetuses. Fibroblasts and neural stem cells (NSCs) were derived from the SCNT fetuses. EGFP was also reactivated in blastocysts after the second SCNT, and induced pluripotent stem cells (iPSCs) were obtained after reprogramming using Yamanaka's factors. The results above indicated that a rabbit reporter system used to monitor the differentiating status of cells was successfully developed.
- Subjects :
- Nuclear Transfer Techniques
Cellular differentiation
Gene Expression
lcsh:Medicine
Bioinformatics
Green fluorescent protein
Animals, Genetically Modified
Mice
Animal Cells
Genes, Reporter
Promoter Regions, Genetic
lcsh:Science
Induced pluripotent stem cell
Cells, Cultured
reproductive and urinary physiology
Plasmid Vectors
Multidisciplinary
Stem Cells
Transfection
Neural stem cell
Cell biology
Transgenic Engineering
embryonic structures
Somatic cell nuclear transfer
Female
Biological Cultures
Rabbits
Cellular Types
Stem cell
Genetic Engineering
Reprogramming
Research Article
Biotechnology
Pluripotent Stem Cells
Induced Pluripotent Stem Cells
Genetic Vectors
Green Fluorescent Proteins
Somatic-Cell Nuclear Transfer
Biology
Research and Analysis Methods
Genetics
Gene Expression and Vector Techniques
Animals
Molecular Biology Techniques
Molecular Biology
Embryonic Stem Cells
Molecular Biology Assays and Analysis Techniques
lcsh:R
fungi
Biology and Life Sciences
Cell Biology
Recombinant Gene Expression Techniques
Cell Cultures
Fibroblasts
Embryo, Mammalian
lcsh:Q
Stem Cell Lines
Octamer Transcription Factor-3
Cloning
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 9
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....fed8836b8fd4c19960ef555008f45255