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  1. KEEP
  2. Theses and Dissertations
  3. Barrett, The Honors College Thesis/Creative Project Collection
  4. RNA-Guided Modification of Synthetic Gene Networks Using CRISPR-Cas Systems
  5. Full metadata

RNA-Guided Modification of Synthetic Gene Networks Using CRISPR-Cas Systems

Full metadata

Description

The ability to edit chromosomal regions is an important tool for the study of gene function and the ability to engineer synthetic gene networks. CRISPR-Cas systems, a bacterial RNA-guided immune system against foreign nucleic acids, have recently been engineered for a plethora of genome engineering and transcriptional regulation applications. Here we employ engineered variants of CRISPR systems in proof-of-principle experiments demonstrating the ability of CRISPR-Cas derived single-DNA-strand cutting enzymes (nickases) to direct host-cell genomic recombination. E.coli is generally regarded as a poorly recombinogenic host with double-stranded DNA breaks being highly lethal. However, CRISPR-guided nickase systems can be easily programmed to make very precise, non-lethal, incisions in genomic regions directing both single reporter gene and larger-scale recombination events deleting up to 36 genes. Genome integrated repetitive elements of variable sizes can be employed as sites for CRISPR induced recombination. We project that single-stranded based editing methodologies can be employed alongside preexisting genome engineering techniques to assist and expedite metabolic engineering and minimalized genome research.

Date Created
2014-05
Contributors
  • Standage-Beier, Kylie S (Author)
  • Wang, Xiao (Thesis director)
  • Haynes, Karmella (Committee member)
  • Barrett, The Honors College (Contributor)
  • School of Life Sciences (Contributor)
Topical Subject
  • Synthetic Biology
  • Recombination
  • E. coli
  • CRISPR
  • Genome Engineering
  • Short Guide RNAs
Resource Type
Text
Extent
22 pages
Language
eng
Copyright Statement
In Copyright
Primary Member of
Barrett, The Honors College Thesis/Creative Project Collection
Series
Academic Year 2013-2014
Handle
https://hdl.handle.net/2286/R.I.22689
Embargo Release Date
Fri, 04/08/2016 - 04:13
Level of coding
minimal
Cataloging Standards
asu1
System Created
  • 2017-10-30 02:50:57
System Modified
  • 2021-08-11 04:09:57
  •     
  • 1 year 7 months ago
Additional Formats
  • OAI Dublin Core
  • MODS XML

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