English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
 
 
DownloadE-Mail
  Morpholino Antisense Oligonucleotide-Mediated Gene Knockdown During Thymocyte Development Reveals Role for Runx3 Transcription Factor in CD4 Silencing During Development of CD4-/CD8+ Thymocytes

Ehlers, M., Laule-Kilian, K., Petter, M., Aldrian, C. J., Grueter, B., Würch, A., et al. (2003). Morpholino Antisense Oligonucleotide-Mediated Gene Knockdown During Thymocyte Development Reveals Role for Runx3 Transcription Factor in CD4 Silencing During Development of CD4-/CD8+ Thymocytes. Journal of Immunology, 171(7), 3594-3604.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002B-94ED-F Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002B-94EE-D
Genre: Journal Article

Files

show Files

Locators

show

Creators

show
hide
 Creators:
Ehlers, Marc1, Author              
Laule-Kilian, Kirsten1, Author              
Petter, Michaela1, Author              
Aldrian, Christine J.1, Author              
Grueter, Baerbel, Author
Würch, Andreas2, Author              
Yoshida, Naomi, Author
Watanabe, Toshio, Author
Satake, Masanobu, Author
Steimle, Viktor1, Author              
Affiliations:
1Spemann Laboratory, Max Planck Institute of Immunobiology and Epigenetics, Max Planck Society, ou_2243655              
2Department of Developmental Immunology, Max Planck Institute of Immunobiology and Epigenetics, Max Planck Society, ou_2243647              

Content

show
hide
Free keywords: -
 Abstract: During thymic T cell development, immature CD4+/CD8+ thymocytes develop into either CD4+/CD8- helper or CD4-/CD8+ CTLs. The molecular mechanisms governing the complex selection and differentiation steps during thymic T cell development are not well understood. Here we developed a novel approach to investigate gene function during thymocyte development. We transfected ex vivo isolated immature thymocytes with gene-specific morpholino antisense oligonucleotides and induced differentiation in cell or organ cultures. A morpholino oligonucleotide specific for CD8α strongly reduces CD8 expression. To our knowledge, this is the first demonstrated gene knockdown by morpholino oligonucleotides in primary lymphocytes. Using this approach, we show here that the transcription factor Runx3 is involved in silencing of CD4 expression during CD8 T cell differentiation. Runx3 protein expression appears late in thymocyte differentiation and is confined to mature CD8 single-positive thymocytes, whereas Runx3 mRNA is transcribed in mature CD4 and CD8 thymocytes. Therefore, Runx3 protein expression is regulated at a post-transcriptional level. The knockdown of Runx3 protein expression through morpholino oligonucleotides inhibited the development of CD4-/CD8+ T cells. Instead, mature cells with a CD4+/CD8+ phenotype accumulated. Potential Runx binding sites were identified in the CD4 gene silencer element, which are bound by Runx protein in EMSAs. Mutagenesis of potential Runx binding sites in the CD4 gene silencer abolished silencing activity in a reporter gene assay, indicating that Runx3 is involved in CD4 gene silencing. The experimental approach developed here should be valuable for the functional analysis of other candidate genes in T cell differentiation.

Details

show
hide
Language(s): eng - English
 Dates: 2003-10-01
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: eDoc: 125038
ISI: 000185548000035
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Journal of Immunology
  Alternative Title : J. Immunol.
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 171 (7) Sequence Number: - Start / End Page: 3594 - 3604 Identifier: ISSN: 0022-1767