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2021

CRISPR

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Full-Text Articles in Life Sciences

Characterization Of The Atpase Activity Of Casding, Christian Cahoon Dec 2021

Characterization Of The Atpase Activity Of Casding, Christian Cahoon

Fall Student Research Symposium 2021

The battle between bacteria and phage has been ongoing for eons. This battle has generated the evolutionary pressure necessary for the development of microbial immune systems. Characterization of these systems has led to the discovery of molecular tools such CRISPR-Cas systems. This system uses a genetic memory of past viral infections coupled with associated proteins to form ribonucleoprotein complexes which seek out and destroy foreign genetic elements. These systems have been repurposed by scientists to create powerful gene editing tools such as Cas9. With such powerful molecular tools being discovered, we have pursued the characterization of a relatively unknown system, …


Determining The Nucleic Acid Binding Affinities Of Crispr-Associated Ding (Casding), Matt Armbrust Dec 2021

Determining The Nucleic Acid Binding Affinities Of Crispr-Associated Ding (Casding), Matt Armbrust

Fall Student Research Symposium 2021

CRISPR-Cas systems are adaptive prokaryotic immune systems that enable host cells to defend against attack from foreign nucleic acids such as phage infections or plasmids. CRISPR-Cas systems are diverse and encompass 2 classes, 6 types, and 33 subtypes. The Type IV-A CRISPR-Cas system from Pseudomonas aeruginosa strain 83 is composed of five different genes (csf1, csf2, csf3, cas6, and dinG). Type IV-A systems are poorly understood, and currently there is little research detailing their biological and biochemical mechanism of immunity. CasDinG, an ancillary protein within the Type IV-A system, is required for an immune response in vivo. However, the role …


Discovering Virally Encoded Proteins That Block Type Iv Crispr Immune Systems, Andrew Williams Dec 2021

Discovering Virally Encoded Proteins That Block Type Iv Crispr Immune Systems, Andrew Williams

Fall Student Research Symposium 2021

Bacteria and the viruses that infect them have been at war from the beginnings of life until today. Due to selective pressure from viral infection, bacteria have evolved various biological defense systems, including CRISPR-Cas systems that use a genetic memory of previous viral encounters to protect against future invasions. However, recently it has been shown that viruses have evolved counter-strategies to evade CRISPR systems. Virally encoded proteins called anti-CRISPRs use a variety of mechanisms to block the activity of CRISPR immune systems in order to infect bacterial cells. The Jackson lab at USU recently showed that a Type IV-A CRISPR-Cas …


Modulation Of Rho Termination : Expectation Vs Reality, Kavya Sri Sai Chegireddy Dec 2021

Modulation Of Rho Termination : Expectation Vs Reality, Kavya Sri Sai Chegireddy

Legacy Theses & Dissertations (2009 - 2024)

Transcription termination factor Rho is an essential protein in many bacteria. Rho is an ATP-dependent RNA helicase that accounts for 20-50% of termination events in Escherichia coli and plays an important role in preventing pervasive transcription that otherwise might be lethal to bacteria. Rho termination usually occurs in the 3’ UTRs and non-coding regions of the DNA, and the actively transcribing RNA is believed to be protected by translating ribosomes from Rho termination since both transcription and translation are a simultaneous process in prokaryotes. However, in reality, Rho terminates transcription of some protein-coding genes, and fails to terminate transcription of …


Evaluating Large Spontaneous Deletions In A Bovine Cell Line Selected For Bovine Viral Diarrhea Virus Resistance, Aspen M. Workman, Michael P. Heaton, Dennis A. Webster, Gregory P. Harhay, Theodore S. Kalbfleisch, Timothy P.L. Smith, Shollie M. Falkenberg, Daniel F. Carlson, Tad S. Sonstegard Nov 2021

Evaluating Large Spontaneous Deletions In A Bovine Cell Line Selected For Bovine Viral Diarrhea Virus Resistance, Aspen M. Workman, Michael P. Heaton, Dennis A. Webster, Gregory P. Harhay, Theodore S. Kalbfleisch, Timothy P.L. Smith, Shollie M. Falkenberg, Daniel F. Carlson, Tad S. Sonstegard

Roman L. Hruska U.S. Meat Animal Research Center: Reports

Bovine viral diarrhea virus’s (BVDV) entry into bovine cells involves attachment of virions to cellular receptors, internalization, and pH-dependent fusion with endosomal membranes. The primary host receptor for BVDV is CD46; however, the complete set of host factors required for virus entry is unknown. The Madin-Darby bovine kidney (MDBK) cell line is susceptible to BVDV infection, while a derivative cell line (CRIB) is resistant at the level of virus entry. We performed complete genome sequencing of each to identify genomic variation underlying the resistant phenotype with the aim of identifying host factors essential for BVDV entry. Three large compound deletions …


Identification Of Secretion Factors That Restructure Neural Tissue In A Model Of Low Grade Glioma, Jackson Ayers Oct 2021

Identification Of Secretion Factors That Restructure Neural Tissue In A Model Of Low Grade Glioma, Jackson Ayers

Holster Scholar Projects

Pediatric Low-grade Gliomas account for up to ⅔ of all pediatric tumors. Despite their low grade classification, they have the ability to mutate farther to a grade 3 or 4 neurological tumor, exponentially decreasing treatment and survival rates. Identifying the specific secretion factors associated with these tumors is extremely beneficial towards possible treatment options. CRISPR/Cas9 constructs were implemented to knock out GDF-15 and CCl-3 secretion factors in the presence of a BRAFV600E mutation before concentrated DNA plasmids were injected and integrated into the brains of mice embryos through In Utero Electroporation (IUE). The brains were subsequently stained through an immunohistochemistry …


Determination Of The Structure, Function, And Mechanism Of Type Iv Crispr-Cas Prokaryotic Defense Systems, Hannah Nicole Taylor Aug 2021

Determination Of The Structure, Function, And Mechanism Of Type Iv Crispr-Cas Prokaryotic Defense Systems, Hannah Nicole Taylor

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Bacteria are under constant threat of invasion by bacteriophage (viruses which infect bacteria). To prevent bacteriophage from entering and overtaking the bacteria, bacteria utilize defense systems to identify and destroy foreign elements. One method of defense is called CRISPR-Cas (Clustered Regularly Interspaced Short Palindromic Repeats – CRISPR-Associated). Many different bacteria and most archaea use CRISPR-Cas systems. There are many diverse types of CRISPR-Cas systems, each of which provides defense in a slightly different way. One such CRISPR-Cas type is called type IV. The type IV CRISPR-Cas system is poorly understood and there are very few studies published on type IV …


Population Genomics Of Invasive Rodents On Islands: Genetic Consequences Of Colonization And Prospects For Localized Synthetic Gene Drive, Kevin P. Oh, Aaron B. Shiels, Laura Shiels, Dimitri V. Blondel, Karl J. Campbell, J. Royden Saah, Alun L. Lloyd, Paul Q. Thomas, Fred Gould, Zaid Abdo, John R. Godwin, Antoinette J. Piaggio May 2021

Population Genomics Of Invasive Rodents On Islands: Genetic Consequences Of Colonization And Prospects For Localized Synthetic Gene Drive, Kevin P. Oh, Aaron B. Shiels, Laura Shiels, Dimitri V. Blondel, Karl J. Campbell, J. Royden Saah, Alun L. Lloyd, Paul Q. Thomas, Fred Gould, Zaid Abdo, John R. Godwin, Antoinette J. Piaggio

USDA Wildlife Services: Staff Publications

Introduced rodent populations pose significant threats worldwide, with particularly severe impacts on islands. Advancements in genome editing have motivated interest in synthetic gene drives that could potentially provide efficient and localized suppression of invasive rodent populations. Application of such technologies will require rigorous population genomic surveys to evaluate population connectivity, taxonomic identification, and to inform design of gene drive localization mechanisms. One proposed approach leverages the predicted shifts in genetic variation that accompany island colonization, wherein founder effects, genetic drift, and island-specific selection are expected to result in locally fixed alleles (LFA) that are variable in neighboring nontarget populations. Engineering …


Genomic Analysis Of Paenibacillus Larvae Bacteriophages, Casey Stamereilers May 2021

Genomic Analysis Of Paenibacillus Larvae Bacteriophages, Casey Stamereilers

UNLV Theses, Dissertations, Professional Papers, and Capstones

American Foulbrood is the most destructive bacterial infection of the honeybee (Apis mellifera) and is caused by the Gram-positive, spore forming bacterium Paenibacillus larvae. Current treatment methods rely on antibiotics, but antibiotics treatments are experiencing a reduction in efficacy due to the recent rise in antibiotic resistant strains of P. larvae. This has been a major catalyst for exploration of alternative treatment methods. Phage therapy is an alternative treatment method that uses viruses that exclusively infect bacteria, known as bacteriophages (phages), to combat bacterial infections. Several experimental studies have shown that phages P. larvae phages are effective at lysing P. …


Crispr/Cas9-Induced Fad2 And Rod1 Mutations Stacked With Fae1 Confer High Oleic Acid Seed Oil In Pennycress (Thlaspi Arvense L.), Brice A. Jarvis, Trevor B. Romsdahl, Michaela G. Mcginn, Tara J. Nazarenus, Edgar B. Cahoon, Kent D. Chapman, John C. Sedbrook Apr 2021

Crispr/Cas9-Induced Fad2 And Rod1 Mutations Stacked With Fae1 Confer High Oleic Acid Seed Oil In Pennycress (Thlaspi Arvense L.), Brice A. Jarvis, Trevor B. Romsdahl, Michaela G. Mcginn, Tara J. Nazarenus, Edgar B. Cahoon, Kent D. Chapman, John C. Sedbrook

Department of Biochemistry: Faculty Publications

Pennycress (Thlaspi arvense L.) is being domesticated as an oilseed cash cover crop to be grown in the off-season throughout temperate regions of the world. With its diploid genome and ease of directed mutagenesis using molecular approaches, pennycress seed oil composition can be rapidly tailored for a plethora of food, feed, oleochemical and fuel uses. Here, we utilized Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology to produce knockout mutations in the FATTY ACID DESATURASE2 (FAD2) and REDUCED OLEATE DESATURATION1 (ROD1) genes to increase oleic acid content. High oleic acid (18:1) oil is valued for its oxidative stability that is …


Crispr And The Transhumanism Movement: Civilian And Military Ethics Reassessment Looming, Jennifer Ha Apr 2021

Crispr And The Transhumanism Movement: Civilian And Military Ethics Reassessment Looming, Jennifer Ha

Student Research Symposium

CRISPR/Cas9 is an emerging biotechnology that allows scientists to edit genes and modify, delete, or correct exact regions of a person’s DNA. This technology could be the future of medicine in eradicating different genetic diseases and disorders and enhancing the quality of life around the world. CRISPR can be used to treat different cancers, blood disorders, AIDS, cystic fibrosis, and a variety of other diseases by going in and changing specific regions of DNA. However, why just eradicate disease when CRISPR can also enhance different characteristics, such as increase metabolism, improve eyesight, or enhance athletic ability? With the potential of …


Reducing Seed Coat Fiber Content And Pod Shatter, And Engineering Medium Chain Fatty Acids-Containing Oil, In The Oilseed Crop Pennycress ( Thlaspi Arvense L. ), Maliheh Esfahanian Mar 2021

Reducing Seed Coat Fiber Content And Pod Shatter, And Engineering Medium Chain Fatty Acids-Containing Oil, In The Oilseed Crop Pennycress ( Thlaspi Arvense L. ), Maliheh Esfahanian

Theses and Dissertations

The overall goal of this thesis was to genetically improve agronomic traits of pennycress (Thlaspi arvense L.; Field Pennycress) and demonstrate the production of value-added designer seed oils to domesticate pennycress and enable its establishment as a new winter annual oilseed/meal/cover crop to be grown in temperate regions of the world. In the U.S. Midwest, pennycress can be double cropped on existing farmland during the time between corn harvest and subsequent planting of soybeans the following spring. Pennycress has the potential to produce 2,000 lbs/acre seeds, which at 33% by weight oil content and 20% protein, would yield 85 gallons/acre …


Characterization Of Crispr Spacer And Protospacer Sequences In Paenibacillus Larvae And Its Bacteriophages, Casey Stamereilers, Simon Wong, Philippos K. Tsourkas Mar 2021

Characterization Of Crispr Spacer And Protospacer Sequences In Paenibacillus Larvae And Its Bacteriophages, Casey Stamereilers, Simon Wong, Philippos K. Tsourkas

Life Sciences Faculty Research

The bacterium Paenibacillus larvae is the causative agent of American foulbrood, the most devastating bacterial disease of honeybees. Because P. larvae is antibiotic resistant, phages that infect it are currently used as alternative treatments. However, the acquisition by P. larvae of CRISPR spacer sequences from the phages could be an obstacle to treatment efforts. We searched nine complete genomes of P. larvae strains and identified 714 CRISPR spacer sequences, of which 384 are unique. Of the four epidemiologically important P. larvae strains, three of these have fewer than 20 spacers, while one strain has over 150 spacers. Of the 384 …