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https://doi.org/10.5562/cca2859

Genome Editing Tools for Functional Analysis of HNF1A and IL6ST Genes

Paula Dobrinić ; University of Zagreb, Faculty of Science, Department of Biology, Division of Molecular Biology, Horvatovac 102a, HR-10000 Zagreb, Croatia
Vanja Tadić ; University of Zagreb, Faculty of Science, Department of Biology, Division of Molecular Biology, Horvatovac 102a, HR-10000 Zagreb, Croatia
Luka Bočkor ; University of Zagreb, Faculty of Science, Department of Biology, Division of Molecular Biology, Horvatovac 102a, HR-10000 Zagreb, Croatia
Dora Markulin ; University of Zagreb, Faculty of Science, Department of Biology, Division of Molecular Biology, Horvatovac 102a, HR-10000 Zagreb, Croatia
Mihaela Tremski ; University of Zagreb, Faculty of Science, Department of Biology, Division of Molecular Biology, Horvatovac 102a, HR-10000 Zagreb, Croatia
Vlatka Zoldoš ; University of Zagreb, Faculty of Science, Department of Biology, Division of Molecular Biology, Horvatovac 102a, HR-10000 Zagreb, Croatia
Aleksandar Vojta ; University of Zagreb, Faculty of Science, Department of Biology, Division of Molecular Biology, Horvatovac 102a, HR-10000 Zagreb, Croatia

Puni tekst: engleski, pdf (1 MB) str. 183-188 preuzimanja: 1.658* citiraj
APA 6th Edition
Dobrinić, P., Tadić, V., Bočkor, L., Markulin, D., Tremski, M., Zoldoš, V. i Vojta, A. (2016). Genome Editing Tools for Functional Analysis of HNF1A and IL6ST Genes. Croatica Chemica Acta, 89 (2), 183-188. https://doi.org/10.5562/cca2859
MLA 8th Edition
Dobrinić, Paula, et al. "Genome Editing Tools for Functional Analysis of HNF1A and IL6ST Genes." Croatica Chemica Acta, vol. 89, br. 2, 2016, str. 183-188. https://doi.org/10.5562/cca2859. Citirano 28.10.2020.
Chicago 17th Edition
Dobrinić, Paula, Vanja Tadić, Luka Bočkor, Dora Markulin, Mihaela Tremski, Vlatka Zoldoš i Aleksandar Vojta. "Genome Editing Tools for Functional Analysis of HNF1A and IL6ST Genes." Croatica Chemica Acta 89, br. 2 (2016): 183-188. https://doi.org/10.5562/cca2859
Harvard
Dobrinić, P., et al. (2016). 'Genome Editing Tools for Functional Analysis of HNF1A and IL6ST Genes', Croatica Chemica Acta, 89(2), str. 183-188. https://doi.org/10.5562/cca2859
Vancouver
Dobrinić P, Tadić V, Bočkor L, Markulin D, Tremski M, Zoldoš V i sur. Genome Editing Tools for Functional Analysis of HNF1A and IL6ST Genes. Croatica Chemica Acta [Internet]. 2016 [pristupljeno 28.10.2020.];89(2):183-188. https://doi.org/10.5562/cca2859
IEEE
P. Dobrinić, et al., "Genome Editing Tools for Functional Analysis of HNF1A and IL6ST Genes", Croatica Chemica Acta, vol.89, br. 2, str. 183-188, 2016. [Online]. https://doi.org/10.5562/cca2859

Sažetak
Genome editing tools, such as TALEN (transcription activator-like effector nuclease) or CRISPR-Cas9 (CRISPR-associated protein-9 nuclease) systems, enable functional studies by targeted gene knockout. They introduce double-stranded breaks (DSBs) into a DNA molecule in a sequence-specific manner, thereby stimulating the error-prone non-homologous end joining repair mechanism, leading to probable gene inactivation when the coding sequence is targeted. Vectors for expression of TALEN and Cas9-based constructs targeting the human IL6ST and HNF1A genes were assembled and tested for their ability to introduce DSBs when transfected into cultured cells using the luciferase assay. The Cas9-based construct targeting the IL6ST gene was shown to be active, while the two TALEN-based constructs did not introduce DSBs above background level. Both the TALEN and the CRISPR-Cas9 constructs targeting the HNF1A gene were found to be active, with the TALEN showing higher activity in a dose-dependent manner. The constructed genome-editing tools can be used for functional analysis of the putative role of HNF1A and IL6ST genes in IgG glycosylation, as shown previously by genome wide association studies.

Creative Commons License This work is licensed under a Creative Commons Attribution 4.0 International License.

Ključne riječi
gene knockout; TALEN; CRISPR; HNF1A; IL6ST; genome editing

Hrčak ID: 161229

URI
https://hrcak.srce.hr/161229

Posjeta: 1.989 *