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[{"key": "dc.contributor.advisor", "value": "Ruotsalainen, Pilvi", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "Jalasvuori, Matti", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.author", "value": "Mikkola, Aapo", "language": "", "element": "contributor", "qualifier": "author", "schema": "dc"}, {"key": "dc.date.accessioned", "value": "2021-03-02T06:49:21Z", "language": null, "element": "date", "qualifier": "accessioned", "schema": "dc"}, {"key": "dc.date.available", "value": "2021-03-02T06:49:21Z", "language": null, "element": "date", "qualifier": "available", "schema": "dc"}, {"key": "dc.date.issued", "value": "2020", "language": "", "element": "date", "qualifier": "issued", "schema": "dc"}, {"key": "dc.identifier.uri", "value": "https://jyx.jyu.fi/handle/123456789/74453", "language": null, "element": "identifier", "qualifier": "uri", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Kasvava antibioottiresistenssi on merkitt\u00e4v\u00e4 ongelma, joka vaikuttaa globaalisti\nterveydenhoitoon, ruoantuotantoon ja taloudelliseen kasvuun. Beta-laktaamien\nantibioottiluokka kattaa noin kaksi kolmasosaa ihmisten k\u00e4ytt\u00e4mist\u00e4 antibiooteista.\nLaajan kirjon beta-laktamaasi (ESBL) -entsyymit mahdollistavat monipuolisen beta-\nlaktaami-resistenssin useassa eri bakteerilajissa. T\u00e4ss\u00e4 tutkimuksessa kloonattiin\nkonjugatiivinen CRISPR-Cas9-plasmidi, joka hy\u00f6kk\u00e4\u00e4 pEC13 ESBL-plasmidia\nvastaan E. Coli-bakteerissa. Tutkimuksen CRISPR-Cas9-plasmidiin liitettiin oriT\n(origin of transfer) -sekvenssi, joka mahdollistaa t\u00e4m\u00e4n kokeellisen plasmidin\nkonjugoimisen k\u00e4ytt\u00e4en toisen pLM2-plasmidin konjugaatiokoneistoa. CRISPR-\nCas9-plasmidiin kohdentamiseksi pEC13-plasmidiin, siihen liitettiin my\u00f6s IncFII\ncrRNA-juoste. T\u00e4m\u00e4n muokatun CRISPR-Cas9-plasmidin tulisi t\u00e4ten pysty\u00e4\nkonjugoitumaan uuteen is\u00e4nt\u00e4soluun pLM2-plasmidin v\u00e4lityksell\u00e4, miss\u00e4 CRISPR-\nCas9-plasmidin Cas9-endonukleaasi leikkaa pEC13-plasmidin DNA-\nkaksoisjuosteen sen IncFII-sekvenssin kohdalta. Leikatun pEC13-plasmidin ei tulisi\nen\u00e4\u00e4 selvit\u00e4 is\u00e4nt\u00e4solussa, mink\u00e4 tulisi johtaa is\u00e4nt\u00e4solun kuolemaan beta-\nlaktaami-maljoilla. Tulokset osoittavat, ett\u00e4 tutkimuksen CRISPR-Cas9 plasmidi\nkonjugoituu uusiin is\u00e4nt\u00e4soluihin ja indusoitu CRISPR-Cas9-plasmidi v\u00e4hent\u00e4\u00e4\nbeta-laktaamilla kasvavien is\u00e4nt\u00e4bakteereiden m\u00e4\u00e4r\u00e4\u00e4 noin kahden kertaluokan\nverran. Kokonaisuutena, t\u00e4t\u00e4 tutkimusta voidaan pit\u00e4\u00e4 onnistuneena konseptin\ntodistuksena, mik\u00e4 n\u00e4ytt\u00e4\u00e4, ett\u00e4 kokeellinen CRISPR-Cas9-plasmidi on mahdollista\nkonjugoida uuteen is\u00e4nt\u00e4bakteeriin, miss\u00e4 sen CRISPR-Cas9-aktiivisuus\nhuomattavasti heikent\u00e4\u00e4 is\u00e4nt\u00e4bakteerien selviytymist\u00e4 beta-laktaami-maljoilla.", "language": "fi", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Emerging antibiotic resistance is one of the major threats to modern healthcare as\nwell as to global food security and economic development. Approximately two-\nthirds of antibiotics administered to humans are \u00df-lactams. The emergence of\nextended-spectrum \u00df-lactamases (ESBLs) in a variety of bacteria confers multi-\nresistance to \u00df-lactams. In this study, an ESBL-harbouring pEC13 plasmid was\ntargeted with a CRISPR-Cas9 plasmid that is delivered to the target E. Coli cells via\nconjugation machinery of the conjugative plasmid pLM2. An anti-ESBL plasmid\nwas cloned, which carries an origin of transfer (oriT) domain for the conjugation\ninitiation along with a gene for CRISPR-Cas9. One specific guiding RNA targeting\nthe IncFII replication initiator gene of the pEC13 plasmid was designed and\nannealed to the anti-ESBL plasmid. In practice, the introduction of a modified anti-\nESBL plasmid through a conjugation channel into an ESBL-harbouring bacterium\nleads to the expression of guiding RNAs that direct the Cas9 endonuclease to cleave\nthe ESBL-plasmid, thus compromising its maintenance in the host. The results show\nthat the induced anti-ESBL plasmid reduces the colony forming unit count of the\nESBL-harbouring host by approximately two orders of magnitude on \u00df-lactam\nplates. The same \u00df-lactam concentration was tested to be lethal without the pEC13\nresistance plasmid. Consequently, this is a viable proof-of-principle study, which\nshows that an anti-ESBL CRISPR-Cas9 plasmid can be introduced into ESBL-\nbacteria via conjugation, and its directed endonuclease activity can substantially\nhinder the survival of those ESBL-bacteria in the presence of lethal \u00df-lactam\nconcentration.", "language": "en", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted by Paivi Vuorio (paelvuor@jyu.fi) on 2021-03-02T06:49:21Z\nNo. of bitstreams: 0", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Made available in DSpace on 2021-03-02T06:49:21Z (GMT). No. of bitstreams: 0\n Previous issue date: 2020", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.format.extent", "value": "53", "language": "", "element": "format", "qualifier": "extent", "schema": "dc"}, {"key": "dc.format.mimetype", "value": "application/pdf", "language": null, "element": "format", "qualifier": "mimetype", "schema": "dc"}, {"key": "dc.language.iso", "value": "eng", "language": null, "element": "language", "qualifier": "iso", "schema": "dc"}, {"key": "dc.rights", "value": "In Copyright", "language": "en", "element": "rights", "qualifier": null, "schema": "dc"}, {"key": "dc.subject.other", "value": "ampicillin", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "beta-lactam", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "CRISPR-Cas9", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "conjugation", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.title", "value": "Abolishment of antibiotic resistance in Escherichia Coli using a conjugative CRISPR-Cas9 plasmid", "language": "", "element": "title", "qualifier": null, "schema": "dc"}, {"key": "dc.type", "value": "master thesis", "language": null, "element": "type", "qualifier": null, "schema": "dc"}, {"key": "dc.identifier.urn", "value": "URN:NBN:fi:jyu-202103021816", "language": "", "element": "identifier", "qualifier": "urn", "schema": "dc"}, {"key": "dc.type.ontasot", "value": "Pro gradu -tutkielma", "language": "fi", "element": "type", "qualifier": "ontasot", "schema": "dc"}, {"key": "dc.type.ontasot", "value": "Master\u2019s thesis", "language": "en", "element": "type", "qualifier": "ontasot", "schema": "dc"}, {"key": "dc.contributor.faculty", "value": "Matemaattis-luonnontieteellinen tiedekunta", "language": "fi", "element": "contributor", "qualifier": "faculty", "schema": "dc"}, {"key": "dc.contributor.faculty", "value": "Faculty of Sciences", "language": "en", "element": "contributor", "qualifier": "faculty", "schema": "dc"}, {"key": "dc.contributor.department", "value": "Bio- ja ymp\u00e4rist\u00f6tieteiden laitos", "language": "fi", "element": "contributor", "qualifier": "department", "schema": "dc"}, {"key": "dc.contributor.department", "value": "Department of Biological and Environmental Science", "language": "en", "element": "contributor", "qualifier": "department", "schema": "dc"}, {"key": "dc.contributor.organization", "value": "Jyv\u00e4skyl\u00e4n yliopisto", "language": "fi", "element": "contributor", "qualifier": "organization", "schema": "dc"}, {"key": "dc.contributor.organization", "value": "University of Jyv\u00e4skyl\u00e4", "language": "en", "element": "contributor", "qualifier": "organization", "schema": "dc"}, {"key": "dc.subject.discipline", "value": "Solu- ja molekyylibiologia", "language": "fi", "element": "subject", "qualifier": "discipline", "schema": "dc"}, {"key": "dc.subject.discipline", "value": "Cell and molecular biology", "language": "en", "element": "subject", "qualifier": "discipline", "schema": "dc"}, {"key": "yvv.contractresearch.funding", "value": "0", "language": "", "element": "contractresearch", "qualifier": "funding", "schema": "yvv"}, {"key": "dc.type.coar", "value": "http://purl.org/coar/resource_type/c_bdcc", "language": null, "element": "type", "qualifier": "coar", "schema": "dc"}, {"key": "dc.rights.accesslevel", "value": "openAccess", "language": null, "element": "rights", "qualifier": "accesslevel", "schema": "dc"}, {"key": "dc.type.publication", "value": "masterThesis", "language": null, "element": "type", "qualifier": "publication", "schema": "dc"}, {"key": "dc.subject.oppiainekoodi", "value": "4013", "language": "", "element": "subject", "qualifier": "oppiainekoodi", "schema": "dc"}, {"key": "dc.subject.yso", "value": "plasmidit", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "antibiootit", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "antibioottiresistenssi", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "kolibakteerit", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "molekyyligenetiikka", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "plasmids", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "antibiotics", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "antibiotic resistance", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "Escherichia coli", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "molecular genetics", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.format.content", "value": "fulltext", "language": null, "element": "format", "qualifier": "content", "schema": "dc"}, {"key": "dc.rights.url", "value": "https://rightsstatements.org/page/InC/1.0/", "language": null, "element": "rights", "qualifier": "url", "schema": "dc"}, {"key": "dc.type.okm", "value": "G2", "language": null, "element": "type", "qualifier": "okm", "schema": "dc"}]
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