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[{"key": "dc.contributor.advisor", "value": "Turkki, Vesa", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "Marjom\u00e4ki, Varpu", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.author", "value": "Lepola, Saana", "language": "", "element": "contributor", "qualifier": "author", "schema": "dc"}, {"key": "dc.date.accessioned", "value": "2019-07-05T08:28:58Z", "language": null, "element": "date", "qualifier": "accessioned", "schema": "dc"}, {"key": "dc.date.available", "value": "2019-07-05T08:28:58Z", "language": null, "element": "date", "qualifier": "available", "schema": "dc"}, {"key": "dc.date.issued", "value": "2019", "language": "", "element": "date", "qualifier": "issued", "schema": "dc"}, {"key": "dc.identifier.uri", "value": "https://jyx.jyu.fi/handle/123456789/64998", "language": null, "element": "identifier", "qualifier": "uri", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Geeniterapiassa sairautta hoidetaan siirt\u00e4m\u00e4ll\u00e4 soluihin siirtogeeni, joka koodaa haluttua proteiinituotetta. Virukset toimivat hyvin\u00e4 geenien kuljettimina eli vektoreina, sill\u00e4 niille on luontaista siirt\u00e4\u00e4 perim\u00e4ns\u00e4 kohdesolun replikoitavaksi. Adenovirus on hyvin yleisesti k\u00e4ytetty geeniterapiavektori, sill\u00e4 sen ominaisuudet tunnetaan hyvin ja se pystyy infektoimaan monenlaisia soluja eri solusyklin vaiheissa. Adenoviruksen rakenneproteiini IX (pIX) sijaitsee heksoniproteiinien v\u00e4liss\u00e4 stabiloiden kapsidin rakennetta. Tutkimukset ovat kuitenkin osoittaneet, ett\u00e4 pIX:ll\u00e4 on mahdollisesti my\u00f6s muunlaisia rooleja adenovirusinfektion aikana. T\u00e4ss\u00e4 tutkimuksessa oli tarkoitus selvitt\u00e4\u00e4, miten kahden genomiltaan erilaisen adenovirusvektorin stabiilisuudet ja infektiivisyydet eroavat toisistaan. Sen lis\u00e4ksi selvitettiin, voidaanko proteiini IX:n m\u00e4\u00e4r\u00e4\u00e4 lis\u00e4t\u00e4 adenoviruksen kapsidissa yliekspressoimalla sit\u00e4 tuottosoluissa ja vaikuttaako m\u00e4\u00e4r\u00e4 adenoviruksen kest\u00e4vyyteen. Molempia vektoreita tuotettiin sek\u00e4 pIX:n yliekpression kanssa ett\u00e4 ilman. Proteiini IX:n lis\u00e4\u00e4misen vaikutusta adenoviruksen kapsidin kest\u00e4vyyteen tutkittiin k\u00e4sittelem\u00e4ll\u00e4 tuotettuja viruksia eri tavoin ja havainnoimalla erot infektiivisyydess\u00e4. Tuotetut virukset puhdistettiin ultrasentrifugoimalla ja dialyysilla. Puhtautta ja pIX:n m\u00e4\u00e4r\u00e4\u00e4 arvioitiin Coomassie-v\u00e4rj\u00e4yksen sek\u00e4 western blottingin avulla. Lis\u00e4ksi virusten tiitterit m\u00e4\u00e4ritettiin ddPCR:n (digital droplet polymerase chain reaction) avulla. Tuotettujen virusten infektiivisyys m\u00e4\u00e4ritettiin infektoimalla tuottosoluja 2 tunnin ajan ja v\u00e4rj\u00e4\u00e4m\u00e4ll\u00e4 solut 48 tunnin kuluttua anti-pIX-vasta-aineella. Tulokset luettiin virtaussytometrilla. Eri vektorien stabiilisuuksissa ei n\u00e4hty tilastollisesti merkitt\u00e4vi\u00e4 eroja yhdess\u00e4k\u00e4\u00e4n k\u00e4sittelykokeessa. Ty\u00f6ss\u00e4 kuitenkin havaittiin, ett\u00e4 vektorit, jotka oli tuotettu pIX:n yliekpression kanssa olivat infektiivisempi\u00e4 kuin ne, jotka oli tuotettu normaalisti tuottosoluissa. Tuloksia voidaan k\u00e4ytt\u00e4\u00e4 hyv\u00e4ksi suunniteltaessa tehokkaampia ja kest\u00e4v\u00e4mpi\u00e4 geeniterapiavektoreita l\u00e4\u00e4keteollisuuden k\u00e4ytt\u00f6\u00f6n.", "language": "fi", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.abstract", "value": "In gene therapy genes are used to treat or cure illnesses. These transgenes are delivered into patient's cells with a special tool, vector. Viruses are often used as vectors because of their natural ability to transfer their genes into target cells. Adenovirus is a very common vector in gene therapy because its features are well known and it can infect many different types of cells in various times in cell cycle. Adenovirus structural protein IX (pIX) is located between hexon proteins stabilizing the structure of adenoviral capsid. Studies indicate that pIX might also have additional roles during adenovirus infection. In this study the object was to find out how the stabilities and infectivities of two different adenoviral vectors differ from each other. The aim was also to find out if the amount of pIX can be increased in adenoviral capsid by overexpressing it in the producer cells and if this has an effect to the stability of the capsid. Both vectors were produced with and without pIX overexpression. The effects of pIX overexpression to stability were studied by treating the viruses in different ways and observing the differences in infectivities. Produced vectors were purified using ultracentrifugation and dialysis. The purity and amount of pIX were estimated using Coomassie staining and Western blot analysis. Virus titers were determined using ddPCR (digital droplet polymerase chain reaction). The infectivities were determined by infecting producer cells for 2 hours and staining the cells after 48 hours with anti-pIX antibody. Results were analyzed using flow cytometry. Although statistically significant differences couldn't be observed in stability studies, the viruses which were produced with overexpression of pIX were more infectious than the ones that had been produced in normal producer cells. These results can be utilized when designing more effective and durable viral vectors for the field of gene therapy.", "language": "en", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted by Miia Hakanen (mihakane@jyu.fi) on 2019-07-05T08:28:58Z\nNo. of bitstreams: 0", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Made available in DSpace on 2019-07-05T08:28:58Z (GMT). No. of bitstreams: 0\n Previous issue date: 2019", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.format.extent", "value": "87", "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": "fin", "language": null, "element": "language", "qualifier": "iso", "schema": "dc"}, {"key": "dc.rights", "value": "In Copyright", "language": "en", "element": "rights", "qualifier": null, "schema": "dc"}, {"key": "dc.title", "value": "Proteiini IX:n yliekspression vaikutus adenovirusvektoreiden stabiilisuuteen ja infektiivisyyteen", "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-201907053584", "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.collaborator", "value": "business", "language": "", "element": "contractresearch", "qualifier": "collaborator", "schema": "yvv"}, {"key": "yvv.contractresearch.funding", "value": "0", "language": "", "element": "contractresearch", "qualifier": "funding", "schema": "yvv"}, {"key": "yvv.contractresearch.initiative", "value": "student", "language": "", "element": "contractresearch", "qualifier": "initiative", "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": "restrictedAccess", "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": "adenovirukset", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "virukset", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "proteiinit", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "geeniterapia", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "geenit", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "tuotto", "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.rights.accessrights", "value": "The author has not given permission to make the work publicly available electronically. Therefore the material can be read only at the archival workstation at Jyv\u00e4skyl\u00e4 University Library (https://kirjasto.jyu.fi/en/workspaces/facilities).", "language": "en", "element": "rights", "qualifier": "accessrights", "schema": "dc"}, {"key": "dc.rights.accessrights", "value": "Tekij\u00e4 ei ole antanut lupaa avoimeen julkaisuun, joten aineisto on luettavissa vain Jyv\u00e4skyl\u00e4n yliopiston kirjaston arkistoty\u00f6semalta. Ks. https://kirjasto.jyu.fi/fi/tyoskentelytilat/laitteet-ja-tilat..", "language": "fi", "element": "rights", "qualifier": "accessrights", "schema": "dc"}, {"key": "dc.type.okm", "value": "G2", "language": null, "element": "type", "qualifier": "okm", "schema": "dc"}]
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