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NRSF/REST is required in vivo for repression of multiple neuronal target genes during embryogenesis

Abstract

The neuron-restrictive silencer factor NRSF (also known as REST and XBR) can silence transcription from neuronal promoters in non-neuronal cell lines, but its function during normal development is unknown. In mice, a targeted mutation of Rest, the gene encoding NRSF, caused derepression of neuron-specific tubulin in a subset of non-neural tissues and embryonic lethality. Mosaic inhibition of NRSF in chicken embryos, using a dominant-negative form of NRSF, also caused derepression of neuronal tubulin, as well as of several other neuronal target genes, in both non-neural tissues and central nervous system neuronal progenitors. These results indicate that NRSF is required to repress neuronal gene expression in vivo, in both extra-neural and undifferentiated neural tissue.

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Figure 1: Targeted disruption of Rest, the gene encoding NRSF.
Figure 2: Early expression of NRSF and morphologic comparison of mutant and wild-type embryos.
Figure 4: Analysis of ßIII tubulin derepression in sections of E9.
Figure 3: The Rest mutation causes derepression of ßIII tubulin in non-neuronal tissues.
Figure 5: Sequence and expression of chick NRSF and inhibition in fibroblasts by dnNRSF.
Figure 6: Derepression of multiple neuronal NRSF target genes in chick embryos injected with dnNRSF into somites.
Figure 7: Derepression and upregulation of neuronal genes in neural crest-derived peripheral glial cells.
Figure 8: Precocious expression of neuronal markers in CNS progenitors of dnNRSF-infected embryos.
Figure 9: Role of NRSF in transcriptional regulation.

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Acknowledgements

We thank R. Behringer and A. Bradley for AB-1 ES cells, SNL 76/7STO cells and FIAU, G. Friedrich for a mouse 129 Sv/Ev genomic library, S.L. Zipursky and B. Wold for helpful discussions and P. Sternberg, K. Zinn, E. Meyerowitz and E. Davidson for their comments on the manuscript. We thank P. Lonnerberg for Ng-CAM cDNA, P. White for help generating CEFs, B. Martinsen and C. Marcelle for advice on RCAS concentration, Shirley Pease, Arti Gaur and members of the TAFCIT staff for expert assistance with chimaera production and mouse maintenance, S. Ou and L. Wang for help with monoclonal antibody production and S. Padilla for technical assistance. This work was supported by a grant from the NIH. A.J.P. is supported by an NIH pre-doctoral training grant. Z.F.C. is an Associate and D.J.A. an Investigator of the Howard Hughes Medical Institute.

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Correspondence to David J. Anderson.

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Chen, ZF., Paquette, A. & Anderson, D. NRSF/REST is required in vivo for repression of multiple neuronal target genes during embryogenesis. Nat Genet 20, 136–142 (1998). https://doi.org/10.1038/2431

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