Naslov
Genome Editing Tools for Functional Analysis of HNF1A and IL6ST Genes

Autori
Dobrinić, Paula ; Tadić, Vanja ; Bočkor, Luka ; Markulin, Dora ; Tremski, Mihaela ; Zoldoš, Vlatka ; Vojta, Aleksandar

Izvornik
Croatica chemica acta (0011-1643) 89 (2016), 2; 183-188

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

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

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.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA