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Directing pluripotent cell differentiation using “diced RNA” in transient transfection

Directing pluripotent cell differentiation using “diced RNA” in transient transfection Embryonic stem (ES) and embryonic carcinoma (EC) cells are pluripotent and have the capacity to differentiate into many cell types. The ability to direct their differentiation should have considerable practical applications. Here, we first report the use of diced short interfering RNAi against Oct4 in a transient approach, to direct differentiation of ES towards the trophectoderm lineage. We then apply this approach to downregulate Smad4 in mouse P19 EC cells. We have found that this leads to an increase in the levels of Pax6 (a neuroectoderm marker), reduction in the levels of Brachyury (a mesoderm marker), and a 3‐fold increase in the number of βIII tubulin‐positive colonies when these cells were allowed to differentiate. This indicates a redirection of cell fate towards the neuroectoderm lineage. Thus, transient RNAi could provide a valuable tool to direct pluripotent cells along specific pathways of differentiation while circumventing permanent genetic changes. genesis 40:157–163, 2004. © 2004 Wiley‐Liss, Inc. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Genesis: the Journal of Genetics and Development Wiley

Directing pluripotent cell differentiation using “diced RNA” in transient transfection

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Publisher
Wiley
Copyright
Copyright © 2004 Wiley Subscription Services, Inc., A Wiley Company
ISSN
1526-954X
eISSN
1526-968X
DOI
10.1002/gene.20078
pmid
15515021
Publisher site
See Article on Publisher Site

Abstract

Embryonic stem (ES) and embryonic carcinoma (EC) cells are pluripotent and have the capacity to differentiate into many cell types. The ability to direct their differentiation should have considerable practical applications. Here, we first report the use of diced short interfering RNAi against Oct4 in a transient approach, to direct differentiation of ES towards the trophectoderm lineage. We then apply this approach to downregulate Smad4 in mouse P19 EC cells. We have found that this leads to an increase in the levels of Pax6 (a neuroectoderm marker), reduction in the levels of Brachyury (a mesoderm marker), and a 3‐fold increase in the number of βIII tubulin‐positive colonies when these cells were allowed to differentiate. This indicates a redirection of cell fate towards the neuroectoderm lineage. Thus, transient RNAi could provide a valuable tool to direct pluripotent cells along specific pathways of differentiation while circumventing permanent genetic changes. genesis 40:157–163, 2004. © 2004 Wiley‐Liss, Inc.

Journal

Genesis: the Journal of Genetics and Development – Wiley

Published: Nov 1, 2004

Keywords: ; ; ; ; ;

References

  • An assessment of the developmental potential of embryonic stem cells in the midgestation mouse embryo
    Beddington, RS; Robertson, EJ
  • Formation of germ‐line chimaeras from embryo‐derived teratocarcinoma cell lines
    Bradley, A
  • A system for stable expression of short interfering RNAs in mammalian cells
    Brummelkamp, TR
  • Distinct regulatory control of the Brachyury gene in axial and non‐axial mesoderm suggests separation of mesoderm lineages early in mouse gastrulation
    Clements, D
  • Establishment in culture of pluripotential cells from mouse embryos
    Evans, MJ; Kaufman, MH
  • Potent and specific genetic interference by double‐stranded RNA in Caenorhabditis elegans
    Fire, A
  • Comment on 'stemness': transcriptional profiling of embryonic and adult stem cells and a stem cell molecular signature
    Fortunel, NO
  • RNA interference by production of short hairpin dsRNA in ES cells, their differentiated derivatives, and in somatic cell lines
    Grabarek, JB
  • RNA interference
    Hannon, GJ
  • Bone morphogenetic proteins in development
    Hogan, BL
  • A stem cell molecular signature
    Ivanova, NB
  • Evidence for involvement of activin A and bone morphogenetic protein 4 in mammalian mesoderm and hematopoietic development
    Johansson, BM; Wiles, MV
  • Retinoic acid induces embryonal carcinoma cells to differentiate into neurons and glial cells
    Jones‐Villeneuve, EM
  • Short hairpin type of dsRNAs that are controlled by tRNA(Val) promoter significantly induce RNAi‐mediated gene silencing in the cytoplasm of human cells
    Kawasaki, H; Taira, K
  • The expression and posttranslational modification of a neuron‐specific beta‐tubulin isotype during chick embryogenesis
    Lee, MK
  • Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells
    Martin, GR
  • Transcriptional control by the TGF‐beta/Smad signaling system
    Massague, J; Wotton, D
  • Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor OCT4
    Nichols, J
  • Self‐renewal of pluripotent embryonic stem cells is mediated via activation of STAT3
    Niwa, H
  • Quantitative expression of Oct‐3/4 defines differentiation, dedifferentiation or self‐renewal of ES cells
    Niwa, H
  • Short hairpin RNAs (shRNAs) induce sequence‐specific silencing in mammalian cells
    Paddison, PJ
  • BMP4 supports self‐renewal of embryonic stem cells by inhibiting mitogen‐activated protein kinase pathways
    Qi, X
  • Stemness: transcriptional profiling of embryonic and adult stem cells
    Ramalho‐Santos, M
  • Primordial germ cell‐derived mouse embryonic germ (EG) cells in vitro resemble undifferentiated stem cells with respect to differentiation capacity and cell cycle distribution
    Rohwedel, J
  • New type of POU domain in germ line‐specific protein Oct‐4
    Scholer, HR
  • Gene expression patterns in human embryonic stem cells and human pluripotent germ cell tumors
    Sperger, JM
  • Mouse embryonic stem cells exhibit indefinite proliferative potential
    Suda, Y
  • Selective reduction of dormant maternal mRNAs in mouse oocytes by RNA interference
    Svoboda, P
  • Gene expression profiling of embryo‐derived stem cells reveals candidate genes associated with pluripotency and lineage specificity
    Tanaka, TS
  • Direct neural fate specification from embryonic stem cells: a primitive mammalian neural stem cell stage acquired through a default mechanism
    Tropepe, V
  • Expanding small RNA interference
    Tuschl, T
  • OCT4 RNA interference induces trophectoderm differentiation in mouse embryonic stem cells
    Velkey, JM; O'Shea, KS
  • Pax‐6, a murine paired box gene, is expressed in the developing CNS
    Walther, C; Gruss, P
  • Specific interference with gene function by double‐stranded RNA in early mouse development
    Wianny, F; Zernicka‐Goetz, M
  • Embryonic stem cell development in a chemically defined medium
    Wiles, MV; Johansson, BM
  • Induction of epidermis and inhibition of neural fate by Bmp‐4
    Wilson, PA; Hemmati‐Brivanlou, A
  • The status of Wnt signalling regulates neural and epidermal fates in the chick embryo
    Wilson, SI
  • Bone morphogenetic protein‐4 is required for mesoderm formation and patterning in the mouse
    Winnier, G
  • BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self‐renewal in collaboration with STAT3
    Ying, QL
  • Consequences of knocking out BMP signalling in the mouse
    Zhao, GQ