Washington University in St. Louis Washington University Open Scholarship Arts & Sciences Electronic Theses and Dissertations Arts & Sciences Summer 8-15-2019 Determining the Genetic Contributions of the Williams Syndrome Critical Region to Behavior Using Mouse Models and Human Genetics Nathan David Kopp Washington University in St. Louis Follow this and additional works at: https://openscholarship.wustl.edu/art_sci_etds Part of the Genetics Commons, and the Neuroscience and Neurobiology Commons Recommended Citation Kopp, Nathan David, "Determining the Genetic Contributions of the Williams Syndrome Critical Region to Behavior Using Mouse Models and Human Genetics" (2019). Arts & Sciences Electronic Theses and Dissertations. 1779. https://openscholarship.wustl.edu/art_sci_etds/1779 This Dissertation is brought to you for free and open access by the Arts & Sciences at Washington University Open Scholarship. It has been accepted for inclusion in Arts & Sciences Electronic Theses and Dissertations by an authorized administrator of Washington University Open Scholarship. For more information, please contact [email protected]. WASHINGTON UNIVERSITY IN ST. LOUIS Division of Biology and Biomedical Sciences Human and Statistical Genetics Dissertation Examination Committee: Joseph Dougherty, Chair Yehuda Ben-Shahar Don Conrad Harrison Gabel Christina Gurnett Beth Kozel Determining the Genetic Contributions of the Williams Syndrome Critical Region to Behavior Using Mouse Models and Human Genetics by Nathan Kopp A dissertation presented to The Graduate School of Washington University in partial fulfillment of the requirements for the degree of Doctor of Philosophy August 2019 St. Louis, Missouri © 2019, Nathan Kopp Table of Contents List of Figures ................................................................................................................................. v List of Tables ................................................................................................................................ vii Acknowledgments .......................................................................................................................... ix Abstract of the Dissertation .......................................................................................................... xii Chapter 1: Introduction ................................................................................................................... 1 1.1 History and description of Williams syndrome ..................................................................... 3 1.2 Genotype-phenotype correlations using human genetics ................................................... 6 1.2.1 Atypical deletions ..................................................................................................................... 7 1.2.2 Human induced pluripotent stem cell (iPSCs) studies ............................................................. 10 1.2.3 Human general population association studies ...................................................................... 14 1.3 Introduction to the general transcription factor 2I family ................................................... 16 1.3.1 General background on the GTF2I family ............................................................................. 17 1.3.2 Mouse models of Gtf2i and Gtf2ird1 ..................................................................................... 23 1.4 Conclusions ......................................................................................................................... 29 Chapter 2: Exome sequencing of 85 Williams Beuren syndrome cases rules out coding variation as a major contributor to remaining variance in social behavior .................................................. 31 2.1 Abstract ............................................................................................................................... 32 2.2 Introduction ......................................................................................................................... 33 2.3 Results ................................................................................................................................. 36 2.3.1 SRS variability in Williams syndrome ....................................................................................... 36 2.3.2 Identification of variants in the Williams syndrome critical region .......................................... 38 2.3.3 Association analyses .................................................................................................................. 39 2.4 Discussion ........................................................................................................................... 43 2.5 Materials and Methods ........................................................................................................ 49 2.6 Acknowledgments ............................................................................................................... 55 2.7 Figures ................................................................................................................................. 56 Chapter 3: The effects of Gtf2ird1 and Gtf2i DNA binding on transcription and behavior supports the important function of the N-terminal end of Gtf2ird1. ............................................. 85 3.1 Abstract ............................................................................................................................... 86 3.2 Introduction ......................................................................................................................... 86 ii 3.3 Results ................................................................................................................................. 88 3.3.1 Gtf2i and Gtf2ird1 bind at active promoters and conserved sites .............................................. 88 3.3.2 Gtf2i and Gtf2ird1 binding sites have distinct features yet overlap at a subset of promoters ... 91 3.3.3 Frameshift mutation in Gtf2ird1 results in truncated protein and affects DNA binding at the Gtf2ird1 promoter ............................................................................................................................... 92 3.3.4 Truncated Gtf2ird1 does not affect binding genome wide ........................................................ 93 3.3.5 Gtf2ird1 frameshift mutation shows mild transcriptional differences ....................................... 94 3.3.6 Frameshift mutation in Gtf2ird1 is sufficient to affect behavior ............................................... 95 3.3.7 Generation of Gtf2i and Gtf2ird1 double mutant ....................................................................... 97 3.3.8 Knocking down both Gtf2i and Gtf2ird1 produces mild transcriptome changes ...................... 98 3.3.9 Double mutants show similar behavioral consequences similar to single Gtf2ird1 mutants ..... 99 3.4 Discussion ......................................................................................................................... 101 3.5 Materials and Methods ...................................................................................................... 105 3.6 Acknowledgements ........................................................................................................... 116 3.7 Figures ............................................................................................................................... 117 Chapter 4: Gtf2i and Gtf2ird1 mutation are not sufficient to reproduce mouse phenotypes caused by the Williams syndrome critical region ................................................................................... 133 4.1 Abstract ............................................................................................................................. 134 4.2 Introduction ....................................................................................................................... 134 4.3 Results ............................................................................................................................... 139 4.3.1 Generation and validation of Gtf2i and Gtf2ird1 loss of function mutation on the same chromosome. ..................................................................................................................................... 139 4.3.2 Gtf2i* mutation is not sufficient to reproduce WSCR-mediated alterations of vocal communication .................................................................................................................................. 143 4.3.3 Gtf2i* mutation is not sufficient to reproduce WSCR-mediated alterations of social behavior ........................................................................................................................................................... 145 4.3.4 Gtf2i* mutation is not sufficient to reproduce WSCR mediated alterations of motor behavior ........................................................................................................................................................... 148 4.3.5 WSCR mutation does not produce robust anxiety-like behaviors ........................................... 151 4.3.6 Gtf2i* mutation is not sufficient to reproduce WSCR mediated alterations of fear conditioning ........................................................................................................................................................... 152 4.3.7 Gtf2i*