University of Illinois Urbana-Champaign

Creating a versatile toolkit for transgene expression using BACS and for dissecting large-scale chromatin organization

Zhao, Binhui

Content Files
Data_A.1.xlsx
Data_C.1.gb
Data_D.1.txt
Data_D.2.txt
ZHAO-DISSERTATION-2019.pdf

Permalink

https://hdl.handle.net/2142/105909

Description

Title
Creating a versatile toolkit for transgene expression using BACS and for dissecting large-scale chromatin organization
Author(s)
Zhao, Binhui
Issue Date
2019-07-09
Director of Research (if dissertation) or Advisor (if thesis)
Belmont, Andrew S
Doctoral Committee Chair(s)
Belmont, Andrew S
Committee Member(s)
  • Prasanth, Supriya
  • Stubbs, Lisa J
  • Freeman, Brian C
  • Zhao, Humin
Department of Study
Cell & Developmental Biology
Discipline
Cell and Developmental Biology
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Date of Ingest
2019-11-26T20:59:34Z
Keyword(s)
  • transgene expression
  • episomes
  • gene amplification
  • human artificial chromosomes
  • DNA synthesis
  • chromatin conformation
  • transcription regulation
  • CRISPR/Cas
Abstract
In eukaryotes, the genetic material, DNA, is highly compacted with histone proteins to form chromatin in interphase nuclei. Both the higher levels of chromatin folding and the spatial organization of the chromatin, referred to as large-scale chromatin organization, have been shown to correlate with transcriptional activity. One example suggesting transcriptional regulation by large-scale chromatin organization is position effects and position effect variegations observed in transgene expressions, which, as well as other epigenetic silencing mechanisms, have been obstacles to achieving predictable and stable transgene expression. Molecular dissections of the determinants regulating large-scale chromatin organization would help to elucidate the real relationship between large-scale chromatin organization and transcriptional regulation, yet are difficult due to the complexity of the mammalian genome. Bacterial artificial chromosomes (BACs), containing 100-300 kb mammalian genomic regions have been shown to recapitulate the expression level and nuclear localization of the corresponding genomic regions, and to protect embedded reporter mini-genes from epigenetic silencing. Here in Chapter 2 we show that BACs could provide a versatile platform for achieving reproducible, stable simultaneous expression of multiple transgenes maintained either as episomes or stably integrated copies. Moreover, in Chapter 3 we show that BACs could be used as a powerful model system for dissecting mechanisms regulating large-scale chromatin organization, by demonstrating distinctive large-scale chromatin conformations formed by BAC transgene arrays and results indicating separation of large-scale chromatin compaction, nuclear localization and transcriptional activities.
Graduation Semester
2019-08
Type of Resource
text
Permalink
http://hdl.handle.net/2142/105909
Copyright and License Information
Copyright 2019 Binhui Zhao

Owning Collections

Dissertations and Theses - Cell and Developmental Biology
Graduate Dissertations and Theses at Illinois PRIMARY
Graduate Theses and Dissertations at Illinois