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Pregled bibliografske jedinice broj: 1209108

Functional validation of GWAS hits associated with IgG glycosylation using CRISPR/dCas9 transient expression system


Miškec, Karlo; Mijakovac, Anika; Frkatović, Azra; Krištić, Jasminka; Vujić, Ana; Lauc, Gordan; Vojta, Aleksandar; Zoldoš, Vlatka
Functional validation of GWAS hits associated with IgG glycosylation using CRISPR/dCas9 transient expression system // Journal of Bioanthropology
Zagreb: Institute for Anthropological Research, 2022. str. 127-127 doi:10.54062/jb (predavanje, nije recenziran, sažetak, znanstveni)


CROSBI ID: 1209108 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Functional validation of GWAS hits associated with IgG glycosylation using CRISPR/dCas9 transient expression system

Autori
Miškec, Karlo ; Mijakovac, Anika ; Frkatović, Azra ; Krištić, Jasminka ; Vujić, Ana ; Lauc, Gordan ; Vojta, Aleksandar ; Zoldoš, Vlatka

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Journal of Bioanthropology / - Zagreb : Institute for Anthropological Research, 2022, 127-127

ISBN
978-953-57695-4-5

Skup
12th ISABS CONFERENCE on Forensic and Anthropological Genetics and Mayo Clinic Lectures in Individualized Medicine

Mjesto i datum
Dubrovnik, Hrvatska, 22.06.2022. - 27.06.2022

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Nije recenziran

Ključne riječi
CRISPR/dCas9, IgG glycosylation, FreeStyle™ 293-F Cells, GWAS

Sažetak
Alternative glycosylation of immunoglobulin G (IgG) Fc region has a crucial role in defining pro- or anti-inflammatory effector function of the antibody. Gene network involved in regulation of IgG glycosylation is still poorly understood because glycoyltransferases and glycosydases are not the major players in this regulatory process. In this study we functionally validated gene loci associated with IgG glycosylation in previous genome-wide association studies (GWAS). We utilized established stably transfected cell line FreeStyle™ 293-F with CRISPR/ dCas9 fusions for direct regulation of genes targeted with specific sgRNAs. This cell system was designed to secrete IgG molecules so that glycans on IgG can be analysed following gene manipulations. We manipulated 22 GWAS hits, grouped according to glycosylation traits such as galactosylation, fucosylation, sialylation and bisecting GlcNAC. Following gene expression manipulations, IgG glycosylation was analysed. Out of seven genes associated with galactosylation, only MANBA, HIVEP2, TNFRSF13B and EEF1A1 showed change of agalactosylated structures when upregulated using dSaCas9-VPR. Out of six hits associated with fucosylation, only upregulation of TBX21 and TBKBP1 showed significant changes in galactosylation, but no change was observed for fucosylated glycan structures. These results suggest that a different cell model might be preferable to validate different GWA hits, such as Lymphoblastoid Cell Line (LCL) which is known to be rich in fucose glycan structures. Out of five loci associated with bisecting GlcNac, upregulation of KDELR2 resulted in an increase in biantennary glycan structures with bisecting GlcNac. The upregulation of DERL2 and RRBP1 resulted in an increase of digalactosylated biantennary glycans. Out of four GWA hits for sialylation, only downregulation of SPPL3 led to hyperglycosylation with concomitant increase in sialylated and galactosylated structures and decrease in agalactosylated glycans, despite the fact that all genes were successfully up- or downregulated. Overall, these results have proven the functional role of several GWA hits which are not glycosyltransferases but are associated with the IgG glycosylation pathway. Ongoing research on LCL cell line might unravel the exact role of these and other GWAS hits.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Biotehnologija u biomedicini (prirodno područje, biomedicina i zdravstvo, biotehničko područje)



POVEZANOST RADA


Projekti:
EK-KF-KK.01.1.1.01.0010 - Znanstveni centar izvrsnosti za personaliziranu brigu o zdravlju (ZCIPersonHealth) (Secenji, Aleksandar, EK - KK.01.1.1.01) ( CroRIS)
EK-EFRR-KK.01.1.1.04.0085 - Genomsko inženjerstvo i genska regulacija u staničnim linijama i modelnim organizacijama tehnologija CRISPR / Cas9 CasMouse (CRISPR/Cas9-CasMouse) (Zoldoš, Vlatka; Gajović, Srećko, EK - Ulaganje u znanost i inovacije) ( CroRIS)

Ustanove:
Prirodoslovno-matematički fakultet, Zagreb,
GENOS d.o.o.

Poveznice na cjeloviti tekst rada:

doi hrcak.srce.hr

Citiraj ovu publikaciju:

Miškec, Karlo; Mijakovac, Anika; Frkatović, Azra; Krištić, Jasminka; Vujić, Ana; Lauc, Gordan; Vojta, Aleksandar; Zoldoš, Vlatka
Functional validation of GWAS hits associated with IgG glycosylation using CRISPR/dCas9 transient expression system // Journal of Bioanthropology
Zagreb: Institute for Anthropological Research, 2022. str. 127-127 doi:10.54062/jb (predavanje, nije recenziran, sažetak, znanstveni)
Miškec, K., Mijakovac, A., Frkatović, A., Krištić, J., Vujić, A., Lauc, G., Vojta, A. & Zoldoš, V. (2022) Functional validation of GWAS hits associated with IgG glycosylation using CRISPR/dCas9 transient expression system. U: Journal of Bioanthropology doi:10.54062/jb.
@article{article, author = {Mi\v{s}kec, Karlo and Mijakovac, Anika and Frkatovi\'{c}, Azra and Kri\v{s}ti\'{c}, Jasminka and Vuji\'{c}, Ana and Lauc, Gordan and Vojta, Aleksandar and Zoldo\v{s}, Vlatka}, year = {2022}, pages = {127-127}, DOI = {10.54062/jb}, keywords = {CRISPR/dCas9, IgG glycosylation, FreeStyle™ 293-F Cells, GWAS}, doi = {10.54062/jb}, isbn = {978-953-57695-4-5}, title = {Functional validation of GWAS hits associated with IgG glycosylation using CRISPR/dCas9 transient expression system}, keyword = {CRISPR/dCas9, IgG glycosylation, FreeStyle™ 293-F Cells, GWAS}, publisher = {Institute for Anthropological Research}, publisherplace = {Dubrovnik, Hrvatska} }
@article{article, author = {Mi\v{s}kec, Karlo and Mijakovac, Anika and Frkatovi\'{c}, Azra and Kri\v{s}ti\'{c}, Jasminka and Vuji\'{c}, Ana and Lauc, Gordan and Vojta, Aleksandar and Zoldo\v{s}, Vlatka}, year = {2022}, pages = {127-127}, DOI = {10.54062/jb}, keywords = {CRISPR/dCas9, IgG glycosylation, FreeStyle™ 293-F Cells, GWAS}, doi = {10.54062/jb}, isbn = {978-953-57695-4-5}, title = {Functional validation of GWAS hits associated with IgG glycosylation using CRISPR/dCas9 transient expression system}, keyword = {CRISPR/dCas9, IgG glycosylation, FreeStyle™ 293-F Cells, GWAS}, publisher = {Institute for Anthropological Research}, publisherplace = {Dubrovnik, Hrvatska} }

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