Porcine OCT4 Reporter System Can Monitor Species-Specific Pluripotency During Somatic Cell Reprogramming.
Seung-Hun KimKwang-Hwan ChoiMingyun LeeDong-Kyung LeeChang-Kyu LeePublished in: Cellular reprogramming (2021)
This study examined the activity and function of pig OCT4 enhancer in porcine reprogramming cells. Dual fluorescent protein reporter systems controlled by the upstream regulatory region of OCT4, which is one of the master regulators for pluripotency, are widely used in studies of the mechanism of pluripotency. We analyzed how this reporter system functions in fibroblast growth factor (FGF)- or leukemia inhibitory factor (LIF)-dependent reprogrammed porcine pluripotent stem cells using the previously established porcine-specific reporter system. Porcine embryonic fibroblasts were coinfected with the pOCT4-ΔPE-eGFP (distal enhancer [DE]-green fluorescent protein [GFP]) and pOCT4-ΔDE-DsRed2 (proximal enhancer [PE]-red fluorescent protein [RFP]) vectors, and GFP and RFP expression were verified during a DOX-dependent reprogramming process. We demonstrated that the porcine OCT4 DE and PE were activated in different expression patterns simultaneously as changes in the expression of pluripotent marker genes during the establishment of porcine-induced pluripotent stem cells (iPSCs). Porcine OCT4 upstream region-derived dual fluorescent protein reporter systems confirmed that porcine iPSCs are in primed state after reprogramming in FGF2- or LIF-containing media. This work demonstrates the applicability of porcine OCT4 upstream region-derived dual fluorescence reporter system, which may be applied to investigations of species-specific pluripotency in porcine-origin cells. These reporter systems may be useful tools for studies of porcine-specific pluripotency, early embryo development, and embryonic stem cells.
Keyphrases
- embryonic stem cells
- binding protein
- crispr cas
- optical coherence tomography
- poor prognosis
- transcription factor
- diabetic retinopathy
- quantum dots
- induced pluripotent stem cells
- stem cells
- induced apoptosis
- cell death
- gene expression
- oxidative stress
- mesenchymal stem cells
- dna methylation
- optic nerve
- amino acid
- minimally invasive
- cell fate
- single molecule
- signaling pathway
- genetic diversity
- endoplasmic reticulum stress
- energy transfer
- cell cycle arrest
- pregnancy outcomes