Biotin tagging of mecp2 in mice reveals contextual insights into the rett syndrome transcriptome

ABSTRACT Mutations in _MECP2_ cause Rett syndrome (RTT), an X-linked neurological disorder characterized by regressive loss of neurodevelopmental milestones and acquired psychomotor

deficits. However, the cellular heterogeneity of the brain impedes an understanding of how _MECP2_ mutations contribute to RTT. Here we developed a Cre-inducible method for

cell-type-specific biotin tagging of MeCP2 in mice. Combining this approach with an allelic series of knock-in mice carrying frequent RTT-associated mutations (encoding T158M and R106W)

enabled the selective profiling of RTT-associated nuclear transcriptomes in excitatory and inhibitory cortical neurons. We found that most gene-expression changes were largely specific to

each RTT-associated mutation and cell type. Lowly expressed cell-type-enriched genes were preferentially disrupted by MeCP2 mutations, with upregulated and downregulated genes reflecting

distinct functional categories. Subcellular RNA analysis in MeCP2-mutant neurons further revealed reductions in the nascent transcription of long genes and uncovered widespread

post-transcriptional compensation at the cellular level. Finally, we overcame X-linked cellular mosaicism in female RTT models and identified distinct gene-expression changes between

neighboring wild-type and mutant neurons, providing contextual insights into RTT etiology that support personalized therapeutic interventions. Access through your institution Buy or

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KNOCKOUT IN HUMAN IPSC-DERIVED GLUTAMATERGIC NEURONS CONFIRMS ITS ROLE IN MTOR SIGNALING AND NEURODEVELOPMENTAL DISORDERS Article 21 July 2023 MITOCHONDRIAL PROTEINS ENCODED BY THE 22Q11.2

NEURODEVELOPMENTAL LOCUS REGULATE NEURAL STEM AND PROGENITOR CELL PROLIFERATION Article Open access 04 October 2023 YY1 MUTATIONS DISRUPT CORTICOGENESIS THROUGH A CELL TYPE SPECIFIC REWIRING

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Download references ACKNOWLEDGEMENTS We would like to thank the IDDRC Mouse Gene Manipulation Core at Children's Hospital Boston (U54HD090255, M. Thompson), the Gene Targeting Core

(P01DK049210, K. Kaestner) and the Transgenic and Chimeric Mouse Facility (J. Richa) at University of Pennsylvania for help in generating transgenic mice, the Flow Cytometry and Cell Sorting

Resource Laboratory (H. Pletcher, W. DeMuth), and the Next Generation Sequencing Core (J. Schug) for technical assistance. B.S.J. is supported by a Cell and Molecular Biology Training Grant

(TG32GM072290) and the UNCF/Merck Graduate Research Dissertation Fellowship. This work is supported by NIH grants K22AI112570 (G.V.), R21AI107067 and R01CA140485 (T.H.K.), R01MH091850 and

R01NS081054 (Z.Z.), and a basic research grant from Rettsyndrome.org (Z.Z.). Z.Z. is a Pew Scholar in the Biomedical Sciences. AUTHOR INFORMATION Author notes * Brian S Johnson, Ying-Tao

Zhao and Maria Fasolino: These authors contributed equally to this work. AUTHORS AND AFFILIATIONS * Department of Genetics, University of Pennsylvania Perelman School of Medicine,

Philadelphia, Pennsylvania, USA Brian S Johnson, Ying-Tao Zhao, Maria Fasolino, Janine M Lamonica, George Georgakilas, Kathleen H Wood, Daniel Bu, Yue Cui, Darren Goffin, Golnaz Vahedi &

 Zhaolan Zhou * Department of Biological Sciences and Center for Systems Biology, University of Texas at Dallas, Richardson, Texas, USA Yoon Jung Kim & Tae Hoon Kim Authors * Brian S

Johnson View author publications You can also search for this author inPubMed Google Scholar * Ying-Tao Zhao View author publications You can also search for this author inPubMed Google

Scholar * Maria Fasolino View author publications You can also search for this author inPubMed Google Scholar * Janine M Lamonica View author publications You can also search for this author

inPubMed Google Scholar * Yoon Jung Kim View author publications You can also search for this author inPubMed Google Scholar * George Georgakilas View author publications You can also

search for this author inPubMed Google Scholar * Kathleen H Wood View author publications You can also search for this author inPubMed Google Scholar * Daniel Bu View author publications You

can also search for this author inPubMed Google Scholar * Yue Cui View author publications You can also search for this author inPubMed Google Scholar * Darren Goffin View author

publications You can also search for this author inPubMed Google Scholar * Golnaz Vahedi View author publications You can also search for this author inPubMed Google Scholar * Tae Hoon Kim

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CONTRIBUTIONS Conceptualization, B.S.J. and Z.Z.; methodology, B.S.J., J.M.L., D.G. and Z.Z.; investigation, B.S.J., Y.-T.Z., M.F., J.M.L., K.H.W., Y.J.K. and D.B.; formal analyses, B.S.J.,

Y.-T.Z., G.G. and T.H.K.; validation, B.S.J., M.F., J.M.L. and G.V.; resources, B.S.J., Y.-T.Z. and Y.C.; data curation, Y.-T.Z.; writing manuscript, B.S.J.; review and editing, B.S.J.,

Y.-T.Z., M.F. and Z.Z.; visualization, B.S.J.; project administration and funding acquisition, Z.Z. CORRESPONDING AUTHOR Correspondence to Zhaolan Zhou. ETHICS DECLARATIONS COMPETING

INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY TEXT AND FIGURES Supplementary Figures 1–13 (PDF 29209 kb) LIFE SCIENCES REPORTING

SUMMARY (PDF 171 KB) SUPPLEMENTARY TABLE 1 Summary of RNA-seq experimental conditions used in this study (XLSX 14 kb) SUPPLEMENTARY TABLE 2 RT-PCR primers used in this study (XLSX 65 kb)

SUPPLEMENTARY TABLE 3 List of HITS-CLIP data sets used for RBP analysis (XLSX 55 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Johnson, B., Zhao,

YT., Fasolino, M. _et al._ Biotin tagging of MeCP2 in mice reveals contextual insights into the Rett syndrome transcriptome. _Nat Med_ 23, 1203–1214 (2017). https://doi.org/10.1038/nm.4406

Download citation * Received: 21 September 2016 * Accepted: 18 August 2017 * Published: 18 September 2017 * Issue Date: 01 October 2017 * DOI: https://doi.org/10.1038/nm.4406 SHARE THIS

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