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

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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…

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.
ContributorsStandage-Beier, Kylie S (Author) / Wang, Xiao (Thesis director) / Haynes, Karmella (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor)
Created2014-05

Direct and indirect recombination rate estimates provide novel insights into the landscape of crossover events in rhesus macaques

Description

Previous recombination rate estimation studies in rhesus macaques have been mostly restricted to a singular approach (e.g., using microsatellite loci). Here, we employ a bilateral method in estimating recombination rates—pedigree-based and linkage-disequilibrium-based—from whole-genome data of rhesus macaques to estimate CO and NCO recombination events and to compare contemporary and historical…

Previous recombination rate estimation studies in rhesus macaques have been mostly restricted to a singular approach (e.g., using microsatellite loci). Here, we employ a bilateral method in estimating recombination rates—pedigree-based and linkage-disequilibrium-based—from whole-genome data of rhesus macaques to estimate CO and NCO recombination events and to compare contemporary and historical rates of recombination.
ContributorsWeiss, Sarah (Author) / Pfeifer, Susanne (Thesis director) / Versoza, Cyril (Committee member) / Barrett, The Honors College (Contributor) / School of Art (Contributor) / School of Life Sciences (Contributor)
Created2023-05

SuppFigures_WeissSarah_Spring_2023.pdf

ContributorsWeiss, Sarah (Author) / Pfeifer, Susanne (Thesis director) / Versoza, Cyril (Committee member) / Barrett, The Honors College (Contributor) / School of Art (Contributor) / School of Life Sciences (Contributor)
Created2023-05

SuppTables_WeissSarah_Spring_2023.pdf

ContributorsWeiss, Sarah (Author) / Pfeifer, Susanne (Thesis director) / Versoza, Cyril (Committee member) / Barrett, The Honors College (Contributor) / School of Art (Contributor) / School of Life Sciences (Contributor)
Created2023-05

WeissSarah_Spring_2023.pdf

ContributorsWeiss, Sarah (Author) / Pfeifer, Susanne (Thesis director) / Versoza, Cyril (Committee member) / Barrett, The Honors College (Contributor) / School of Art (Contributor) / School of Life Sciences (Contributor)
Created2023-05