CRISPR-on system for the activation of the endogenous human INS gene
| Autor: | L Grosembacher, A Mutto, F Pereyra-Bonnet, P Argibay, Carla Alejandra Gimenez, M Ielpi |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Therapeutic gene modulation Epigenomics Transcriptional Activation CRISPRS Otras Ciencias Biológicas Cell Culture Techniques ESC Biology Gene dosage Methylation Ciencias Biológicas 03 medical and health sciences Mice 0302 clinical medicine Epigenetics of physical exercise Genetics CRISPR Animals Humans Insulin Clustered Regularly Interspaced Short Palindromic Repeats Epigenetics Promoter Regions Genetic Molecular Biology Regulation of gene expression Gene targeting INSULIN 030104 developmental biology HEK293 Cells Gene Expression Regulation RNA Molecular Medicine Genetic Engineering CIENCIAS NATURALES Y EXACTAS 030217 neurology & neurosurgery |
| Zdroj: | Gene therapy. 23(6) |
| ISSN: | 1476-5462 |
| Popis: | Advances in the field of epigenetics have allowed the design of new therapeutic strategies to address complex diseases such as type 1 diabetes (T1D). Clustered regularly interspaced short palindromic repeats (CRISPR)-on is a novel and powerful RNA-guided transcriptional activator system that can turn on specific gene expression; however, it remains unclear whether this system can be widely used or whether its use will be restricted depending on cell types, methylation promoter statuses or the capacity to modulate chromatin state. Our results revealed that the CRISPR-on system fused with transcriptional activators (dCas9-VP160) activated endogenous human INS, which is a silenced gene with a fully methylated promoter. Similarly, we observed a synergistic effect on gene activation when multiple single guide RNAs were used, and the transcriptional activation was maintained until day 21. Regarding the epigenetic profile, the targeted promoter gene did not exhibit alteration in its methylation status but rather exhibited altered levels of H3K9ac following treatment. Importantly, we showed that dCas9-VP160 acts on patients' cells in vitro, particularly the fibroblasts of patients with T1D. Fil: Giménez, Carla Alejandra. Hospital Italiano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ielpi, Marcelo. Hospital Italiano; Argentina Fil: Mutto, Adrián Angel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Grosembacher, Luis. Hospital Italiano; Argentina Fil: Argibay, Pablo. Hospital Italiano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Pereyra Bonnet, Federico Alberto. Hospital Italiano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina |
| Databáze: | OpenAIRE |
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