NANOG reporter cell lines generated by gene targeting in human embryonic stem cells.
Pluripotency and self-renewal of human embryonic stem cells (hESCs) is mediated by a complex interplay between extra- and intracellular signaling pathways, which regulate the expression of pluripotency-specific transcription factors. The homeodomain transcription factor NANOG plays a central role in...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2010-09-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC2932718?pdf=render |
| _version_ | 1811281910773579776 |
|---|---|
| author | Yvonne Fischer Elvira Ganic Jacqueline Ameri Xiaojie Xian Martina Johannesson Henrik Semb |
| author_facet | Yvonne Fischer Elvira Ganic Jacqueline Ameri Xiaojie Xian Martina Johannesson Henrik Semb |
| author_sort | Yvonne Fischer |
| collection | DOAJ |
| description | Pluripotency and self-renewal of human embryonic stem cells (hESCs) is mediated by a complex interplay between extra- and intracellular signaling pathways, which regulate the expression of pluripotency-specific transcription factors. The homeodomain transcription factor NANOG plays a central role in maintaining hESC pluripotency, but the precise role and regulation of NANOG are not well defined.To facilitate the study of NANOG expression and regulation in viable hESC cultures, we generated fluorescent NANOG reporter cell lines by gene targeting in hESCs. In these reporter lines, the fluorescent reporter gene was co-expressed with endogenous NANOG and responded to experimental induction or repression of the NANOG promoter with appropriate changes in expression levels. Furthermore, NANOG reporter lines facilitated the separation of hESC populations based on NANOG expression levels and their subsequent characterization. Gene expression arrays on isolated hESC subpopulations revealed genes with differential expression in NANOG(high) and NANOG(low) hESCs, providing candidates for NANOG downstream targets hESCs.The newly derived NANOG reporter hESC lines present novel tools to visualize NANOG expression in viable hESCs. In future applications, these reporter lines can be used to elucidate the function and regulation of NANOG in pluripotent hESCs. |
| first_indexed | 2024-04-13T01:42:06Z |
| format | Article |
| id | doaj.art-982d42a0745949a69e9cc6d68f204847 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-13T01:42:06Z |
| publishDate | 2010-09-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-982d42a0745949a69e9cc6d68f2048472022-12-22T03:08:08ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-09-015910.1371/journal.pone.0012533NANOG reporter cell lines generated by gene targeting in human embryonic stem cells.Yvonne FischerElvira GanicJacqueline AmeriXiaojie XianMartina JohannessonHenrik SembPluripotency and self-renewal of human embryonic stem cells (hESCs) is mediated by a complex interplay between extra- and intracellular signaling pathways, which regulate the expression of pluripotency-specific transcription factors. The homeodomain transcription factor NANOG plays a central role in maintaining hESC pluripotency, but the precise role and regulation of NANOG are not well defined.To facilitate the study of NANOG expression and regulation in viable hESC cultures, we generated fluorescent NANOG reporter cell lines by gene targeting in hESCs. In these reporter lines, the fluorescent reporter gene was co-expressed with endogenous NANOG and responded to experimental induction or repression of the NANOG promoter with appropriate changes in expression levels. Furthermore, NANOG reporter lines facilitated the separation of hESC populations based on NANOG expression levels and their subsequent characterization. Gene expression arrays on isolated hESC subpopulations revealed genes with differential expression in NANOG(high) and NANOG(low) hESCs, providing candidates for NANOG downstream targets hESCs.The newly derived NANOG reporter hESC lines present novel tools to visualize NANOG expression in viable hESCs. In future applications, these reporter lines can be used to elucidate the function and regulation of NANOG in pluripotent hESCs.http://europepmc.org/articles/PMC2932718?pdf=render |
| spellingShingle | Yvonne Fischer Elvira Ganic Jacqueline Ameri Xiaojie Xian Martina Johannesson Henrik Semb NANOG reporter cell lines generated by gene targeting in human embryonic stem cells. PLoS ONE |
| title | NANOG reporter cell lines generated by gene targeting in human embryonic stem cells. |
| title_full | NANOG reporter cell lines generated by gene targeting in human embryonic stem cells. |
| title_fullStr | NANOG reporter cell lines generated by gene targeting in human embryonic stem cells. |
| title_full_unstemmed | NANOG reporter cell lines generated by gene targeting in human embryonic stem cells. |
| title_short | NANOG reporter cell lines generated by gene targeting in human embryonic stem cells. |
| title_sort | nanog reporter cell lines generated by gene targeting in human embryonic stem cells |
| url | http://europepmc.org/articles/PMC2932718?pdf=render |
| work_keys_str_mv | AT yvonnefischer nanogreportercelllinesgeneratedbygenetargetinginhumanembryonicstemcells AT elviraganic nanogreportercelllinesgeneratedbygenetargetinginhumanembryonicstemcells AT jacquelineameri nanogreportercelllinesgeneratedbygenetargetinginhumanembryonicstemcells AT xiaojiexian nanogreportercelllinesgeneratedbygenetargetinginhumanembryonicstemcells AT martinajohannesson nanogreportercelllinesgeneratedbygenetargetinginhumanembryonicstemcells AT henriksemb nanogreportercelllinesgeneratedbygenetargetinginhumanembryonicstemcells |