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  4. Developing engineered polymerases for practical applications in synthetic biology
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Developing engineered polymerases for practical applications in synthetic biology

Full metadata

Description

Advances in chemical synthesis have enabled new lines of research with unnatural genetic polymers whose modified bases or sugar-phosphate backbones have potential therapeutic and biotechnological applications. Maximizing the potential of these synthetic genetic systems requires inventing new molecular biology tools that can both generate and faithfully replicate unnatural polymers of significant length. Threose nucleic acid (TNA) has received significant attention as a complete replication system has been developed by engineering natural polymerases to broaden their substrate specificity. The system, however, suffers from a high mutational load reducing its utility. This thesis will cover the development of two new polymerases capable of transcribing and reverse transcribing TNA polymers with high efficiency and fidelity. The polymerases are identified using a new strategy wherein gain-of-function mutations are sampled in homologous protein architectures leading to subtle optimization of protein function. The new replication system has a fidelity that supports the propagation of genetic information enabling in vitro selection of functional TNA molecules. TNA aptamers to human alpha-thrombin are identified and demonstrated to have superior stability compared to DNA and RNA in biologically relevant conditions. This is the first demonstration that functional TNA molecules have potential in biotechnology and molecular medicine.

Date Created
2015
Contributors
  • Dunn, Matthew Ryan (Author)
  • Chaput, John C (Thesis advisor)
  • LaBaer, Joshua (Committee member)
  • Lake, Douglas (Committee member)
  • Mangone, Marco (Committee member)
  • Arizona State University (Publisher)
Topical Subject
  • Molecular Biology
  • Biochemistry
  • Polymerase Engineering
  • synthetic genetics
  • Threose Nucleic Acid
  • Xenonucleic Acids
  • Polymers
  • Synthetic Biology
  • DNA polymerases
  • RNA polymerases
  • Nucleic Acids
Resource Type
Text
Genre
Doctoral Dissertation
Academic theses
Extent
ix, 113 pages : illustrations (some color)
Language
eng
Copyright Statement
In Copyright
Reuse Permissions
All Rights Reserved
Primary Member of
ASU Electronic Theses and Dissertations
Peer-reviewed
No
Open Access
No
Handle
https://hdl.handle.net/2286/R.I.35429
Statement of Responsibility
by Matthew Ryan Dunn
Description Source
Retrieved on Dec. 8, 2015
Level of coding
full
Note
Partial requirement for: Ph.D., Arizona State University, 2015
Note type
thesis
Includes bibliographical references (pages 99-113)
Note type
bibliography
Field of study: Molecular and cellular biology
System Created
  • 2015-10-01 08:01:19
System Modified
  • 2021-08-30 01:26:49
  •     
  • 1 year 6 months ago
Additional Formats
  • OAI Dublin Core
  • MODS XML

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