TY  - DATA
AU  - Đorđević, Marija
AU  - Stepper, Peter
AU  - Feuerstein-Akgoz, Clarissa
AU  - Gerhauser, Clarissa
AU  - Paunović, Verica
AU  - Tolić, Anja
AU  - Rajić, Jovana
AU  - Dinić, Svetlana
AU  - Uskoković, Aleksandra
AU  - Grdović, Nevena
AU  - Mihailović, Mirjana
AU  - Jurkowska, Renata
AU  - Jurkowski, Tomasz
AU  - Arambašić Jovanović, Jelena
AU  - Vidaković, Melita
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5669
AB  - Figure 1. UCSC genome browser views of murine Arx gene’s DNA methylation sites in the pancreas and other tissues. If compared with other tissues, Arx gene in pancreatic tissue (young and old) exhibits low DNA methylation. CpG island was shown as a green box. Table 1. Targeted sequences for sgRNAs. Table 2. Primers used for RT-qPCR. Table 3. Primers used for HRM. Table 4. Touchdown PCR program for amplification of bisulfite converted DNA, starting at 55 °C. Table 5. Primers for NGS library preparation. Table 6. Antibodies used for Immunoblot analysis (IBA) and Immunocytochemistry (ICC). Table 7. Primers for PCR reaction after ChIP.
PB  - Frontiers Media S.A.
T2  - Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms
T1  - Supplementary Material for Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska RZ, Jurkowski TP, Jovanović JA, Vidaković M. EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. Front Endocrinol (Lausanne). 2023;14:1134478.
VL  - 14
SP  - 1134478
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5669
ER  - 

@misc{
author = "Đorđević, Marija and Stepper, Peter and Feuerstein-Akgoz, Clarissa and Gerhauser, Clarissa and Paunović, Verica and Tolić, Anja and Rajić, Jovana and Dinić, Svetlana and Uskoković, Aleksandra and Grdović, Nevena and Mihailović, Mirjana and Jurkowska, Renata and Jurkowski, Tomasz and Arambašić Jovanović, Jelena and Vidaković, Melita",
year = "2023",
abstract = "Figure 1. UCSC genome browser views of murine Arx gene’s DNA methylation sites in the pancreas and other tissues. If compared with other tissues, Arx gene in pancreatic tissue (young and old) exhibits low DNA methylation. CpG island was shown as a green box. Table 1. Targeted sequences for sgRNAs. Table 2. Primers used for RT-qPCR. Table 3. Primers used for HRM. Table 4. Touchdown PCR program for amplification of bisulfite converted DNA, starting at 55 °C. Table 5. Primers for NGS library preparation. Table 6. Antibodies used for Immunoblot analysis (IBA) and Immunocytochemistry (ICC). Table 7. Primers for PCR reaction after ChIP.",
publisher = "Frontiers Media S.A.",
journal = "Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms",
title = "Supplementary Material for Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska RZ, Jurkowski TP, Jovanović JA, Vidaković M. EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. Front Endocrinol (Lausanne). 2023;14:1134478.",
volume = "14",
pages = "1134478",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5669"
}

Đorđević, M., Stepper, P., Feuerstein-Akgoz, C., Gerhauser, C., Paunović, V., Tolić, A., Rajić, J., Dinić, S., Uskoković, A., Grdović, N., Mihailović, M., Jurkowska, R., Jurkowski, T., Arambašić Jovanović, J.,& Vidaković, M.. (2023). Supplementary Material for Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska RZ, Jurkowski TP, Jovanović JA, Vidaković M. EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. Front Endocrinol (Lausanne). 2023;14:1134478.. in Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms
Frontiers Media S.A.., 14, 1134478.
https://hdl.handle.net/21.15107/rcub_ibiss_5669

Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska R, Jurkowski T, Arambašić Jovanović J, Vidaković M. Supplementary Material for Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska RZ, Jurkowski TP, Jovanović JA, Vidaković M. EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. Front Endocrinol (Lausanne). 2023;14:1134478.. in Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms. 2023;14:1134478.
https://hdl.handle.net/21.15107/rcub_ibiss_5669 .

Đorđević, Marija, Stepper, Peter, Feuerstein-Akgoz, Clarissa, Gerhauser, Clarissa, Paunović, Verica, Tolić, Anja, Rajić, Jovana, Dinić, Svetlana, Uskoković, Aleksandra, Grdović, Nevena, Mihailović, Mirjana, Jurkowska, Renata, Jurkowski, Tomasz, Arambašić Jovanović, Jelena, Vidaković, Melita, "Supplementary Material for Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska RZ, Jurkowski TP, Jovanović JA, Vidaković M. EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. Front Endocrinol (Lausanne). 2023;14:1134478." in Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms, 14 (2023):1134478,
https://hdl.handle.net/21.15107/rcub_ibiss_5669 .

TY  - JOUR
AU  - Đorđević, Marija
AU  - Stepper, Peter
AU  - Feuerstein-Akgoz, Clarissa
AU  - Gerhauser, Clarissa
AU  - Paunović, Verica
AU  - Tolić, Anja
AU  - Rajić, Jovana
AU  - Dinić, Svetlana
AU  - Uskoković, Aleksandra
AU  - Grdović, Nevena
AU  - Mihailović, Mirjana
AU  - Jurkowska, Renata
AU  - Jurkowski, Tomasz
AU  - Arambašić Jovanović, Jelena
AU  - Vidaković, Melita
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5507
AB  - Introduction: Beta cell dysfunction by loss of beta cell identity, dedifferentiation, and the presence of polyhormonal cells are main characteristics of diabetes. The straightforward strategy for curing diabetes implies reestablishment of pancreatic beta cell function by beta cell replacement therapy. Aristaless-related homeobox (Arx) gene encodes protein which plays an important role in the development of pancreatic alpha cells and is a main target for changing alpha cell identity.

Results: In this study we used CRISPR/dCas9-based epigenetic tools for targeted hypermethylation of Arx gene promoter and its subsequent suppression in mouse pancreatic αTC1-6 cell line. Bisulfite sequencing and methylation profiling revealed that the dCas9-Dnmt3a3L-KRAB single chain fusion constructs (EpiCRISPR) was the most efficient. Epigenetic silencing of Arx expression was accompanied by an increase in transcription of the insulin gene (Ins2) mRNA on 5th and 7th post-transfection day, quantified by both RT-qPCR and RNA-seq. Insulin production and secretion was determined by immunocytochemistry and ELISA assay, respectively. Eventually, we were able to induce switch of approximately 1% of transiently transfected cells which were able to produce 35% more insulin than Mock transfected alpha cells.

Conclusion: In conclusion, we successfully triggered a direct, transient switch of pancreatic alpha to insulin-producing cells opening a future research on promising therapeutic avenue for diabetes management.
PB  - Frontiers Media S.A.
T2  - Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms
T1  - EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells
VL  - 14
DO  - 10.3389/fendo.2023.1134478
SP  - 1134478
ER  - 

@article{
author = "Đorđević, Marija and Stepper, Peter and Feuerstein-Akgoz, Clarissa and Gerhauser, Clarissa and Paunović, Verica and Tolić, Anja and Rajić, Jovana and Dinić, Svetlana and Uskoković, Aleksandra and Grdović, Nevena and Mihailović, Mirjana and Jurkowska, Renata and Jurkowski, Tomasz and Arambašić Jovanović, Jelena and Vidaković, Melita",
year = "2023",
abstract = "Introduction: Beta cell dysfunction by loss of beta cell identity, dedifferentiation, and the presence of polyhormonal cells are main characteristics of diabetes. The straightforward strategy for curing diabetes implies reestablishment of pancreatic beta cell function by beta cell replacement therapy. Aristaless-related homeobox (Arx) gene encodes protein which plays an important role in the development of pancreatic alpha cells and is a main target for changing alpha cell identity.

Results: In this study we used CRISPR/dCas9-based epigenetic tools for targeted hypermethylation of Arx gene promoter and its subsequent suppression in mouse pancreatic αTC1-6 cell line. Bisulfite sequencing and methylation profiling revealed that the dCas9-Dnmt3a3L-KRAB single chain fusion constructs (EpiCRISPR) was the most efficient. Epigenetic silencing of Arx expression was accompanied by an increase in transcription of the insulin gene (Ins2) mRNA on 5th and 7th post-transfection day, quantified by both RT-qPCR and RNA-seq. Insulin production and secretion was determined by immunocytochemistry and ELISA assay, respectively. Eventually, we were able to induce switch of approximately 1% of transiently transfected cells which were able to produce 35% more insulin than Mock transfected alpha cells.

Conclusion: In conclusion, we successfully triggered a direct, transient switch of pancreatic alpha to insulin-producing cells opening a future research on promising therapeutic avenue for diabetes management.",
publisher = "Frontiers Media S.A.",
journal = "Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms",
title = "EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells",
volume = "14",
doi = "10.3389/fendo.2023.1134478",
pages = "1134478"
}

Đorđević, M., Stepper, P., Feuerstein-Akgoz, C., Gerhauser, C., Paunović, V., Tolić, A., Rajić, J., Dinić, S., Uskoković, A., Grdović, N., Mihailović, M., Jurkowska, R., Jurkowski, T., Arambašić Jovanović, J.,& Vidaković, M.. (2023). EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. in Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms
Frontiers Media S.A.., 14, 1134478.
https://doi.org/10.3389/fendo.2023.1134478

Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska R, Jurkowski T, Arambašić Jovanović J, Vidaković M. EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. in Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms. 2023;14:1134478.
doi:10.3389/fendo.2023.1134478 .

Đorđević, Marija, Stepper, Peter, Feuerstein-Akgoz, Clarissa, Gerhauser, Clarissa, Paunović, Verica, Tolić, Anja, Rajić, Jovana, Dinić, Svetlana, Uskoković, Aleksandra, Grdović, Nevena, Mihailović, Mirjana, Jurkowska, Renata, Jurkowski, Tomasz, Arambašić Jovanović, Jelena, Vidaković, Melita, "EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells" in Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms, 14 (2023):1134478,
https://doi.org/10.3389/fendo.2023.1134478 . .