자료유형  

 학위논문

Control Number  

0017161505

International Standard Book Number  

9798382232881

Dewey Decimal Classification Number  

574

Main Entry-Personal Name  

Hinks, Michaela Marie.

Publication, Distribution, etc. (Imprint  

[S.l.] : Stanford University., 2024

Publication, Distribution, etc. (Imprint  

Ann Arbor : ProQuest Dissertations & Theses, 2024

Physical Description  

111 p.

General Note  

Source: Dissertations Abstracts International, Volume: 85-11, Section: B.

General Note  

Advisor: Bintu, Lacramioara;Greenleaf, William.

Dissertation Note  

Thesis (Ph.D.)--Stanford University, 2024.

Summary, Etc.  

요약The binding of multiple transcription factors (TFs) to genomic enhancers activates gene expression in mammalian cells. However, the molecular details that link enhancer sequence to TF binding, promoter state, and gene expression levels remain opaque. We applied single-molecule footprinting (SMF) to measure the simultaneous occupancy of TFs, nucleosomes, and components of the transcription machinery on engineered enhancer/promoter constructs with variable numbers of TF binding sites for both a synthetic and an endogenous TF. We find that activation domains enhance a TF's capacity to compete with nucleosomes for binding to DNA in a BAF-dependent manner, TF binding on nucleosome-free DNA is consistent with independent binding between TFs, and average TF occupancy linearly contributes to promoter activation rates. We also decompose TF strength into separable binding and activation terms, which can be tuned and perturbed independently. Finally, we develop thermodynamic and kinetic models that quantitatively predict both the binding microstates observed at the enhancer and subsequent time-dependent gene expression. Chapters 2-4 provide a template for quantitative dissection of distinct contributors to gene activation, including the activity of chromatin remodelers, TF activation domains, chromatin acetylation, TF concentration, TF binding affinity, and TF binding site configuration. Chapter 5 demonstrates how novel large serine recombinases can facilitate highly controlled and fast parallel DNA reporter assays by integrating linear DNA library members into landing pads. Chapter 6 characterizes how transcriptional run-on between two tandem genes can lead to unexpected gene expression dynamics in synthetic gene systems. Taken together, this thesis provides novel biological insights and tools to advance understanding and engineering of gene expression in human cells.

Subject Added Entry-Topical Term  

Cellular biology.

Subject Added Entry-Topical Term  

Genetics.

Index Term-Uncontrolled  

Transcription factors

Index Term-Uncontrolled  

Single-molecule footprinting

Index Term-Uncontrolled  

Gene expression

Added Entry-Corporate Name  

Stanford University.

Host Item Entry  

Dissertations Abstracts International. 85-11B.

Electronic Location and Access  

로그인을 한후 보실 수 있는 자료입니다.

Control Number  

joongbu:657713