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.
Details
Title
- RNA-Guided Modification of Synthetic Gene Networks Using CRISPR-Cas Systems
Contributors
- Standage-Beier, Kylie S (Author)
- Wang, Xiao (Thesis director)
- Haynes, Karmella (Committee member)
- Barrett, The Honors College (Contributor)
- School of Life Sciences (Contributor)
Date Created
The date the item was original created (prior to any relationship with the ASU Digital Repositories.)
2014-05
Resource Type
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