magistrsko delo
Meta Kodrič (Avtor), Duško Lainšček (Mentor), Marko Dolinar (Član komisije za zagovor), Gregor Gunčar (Član komisije za zagovor), Ajda Taler-Verčič (Komentor)

Povzetek

Uporaba sistema CCExo-streptavidin za tarčno integracijo zapisa za CD19-CAR v genom limfocitov T

Ključne besede

hematološka rakava obolenja;celična terapija CAR-T;tehnologija CRISPR/Cas9;sistem CCExo;streptavidin;magistrska dela;

Podatki

Jezik: Slovenski jezik
Leto izida:
Tipologija: 2.09 - Magistrsko delo
Organizacija: UL FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Založnik: [M. Kodrič]
UDK: 577.27:616.15(043.2)
COBISS: 174379779 Povezava se bo odprla v novem oknu
Št. ogledov: 186
Št. prenosov: 0
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Angleški jezik
Sekundarni naslov: Use of the CCExo-streptavidin system for targeted integration of the CD19-CAR transcript into the genome of T lymphocytes
Sekundarni povzetek: CAR-T cell therapy is a form of cancer immunotherapy that has proven itself as an effective cure for treating various haematological malignancies. Generally, viral transduction is used for the preparation of therapeutic cells, yet due to its numerous disadvantages various non-viral methods for CAR-T cell preparation are being investigated, which would be safer, cheaper, and more effective. Hence, as part of the master’s thesis, we prepared an improved version of the CRISPR/Cas9 technology, named CCExo-streptavidin, which enables efficient integration of genes at the target site in the cell genome and thus the preparation of therapeutic cells. The components of the CCExo-streptavidin system were designed and prepared so that they enabled insertion of the second generation anti-CD19 CAR receptor coding region into the cell genome in place of the TRAC gene, which was knocked out. The effectiveness of the CCExo streptavidin system was demonstrated in vivo on the CD4+ T lymphocyte cell line Jurkat and on human primary CD3+ T lymphocytes. It was shown as a higher expression of the CAR receptor on the surface of the treated cells, a higher concentration of IL-2 in the co-culture of T lymphocytes and target cancer cells Raji-fLUC, and a higher cytotoxic activity of the therapeutic CD8+ cells compared to the classic CRISPR/Cas9 system. Further optimization of the CCExo-streptavidin system enabled us to determine the optimal concentration of the components for electroporation of Jurkat cells, yet the optimization can be further improved and carried out additionally for the electroporation of human primary T lymphocytes. Lastly, we aspired to insert the CAR transgene also in the TRBC1 locus, but with little success.
Sekundarne ključne besede: CAR-T;cancer immunotherapy;CRISPR/Cas9;CCExo;streptavidin;Imunoterapija;Univerzitetna in visokošolska dela;
Vrsta dela (COBISS): Magistrsko delo/naloga
Študijski program: 1000377
Konec prepovedi (OpenAIRE): 2024-10-26
Komentar na gradivo: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, smer Biokemija
Strani: 79 str.
ID: 20605151