Waking up dormant tumor suppressor genes with zinc fingers, TALEs and the CRISPR/dCas9 system
The aberrant epigenetic silencing of tumor suppressor genes (TSGs) plays a major role during carcinogenesis and regaining these dormant functions by engineering of sequence-specific epigenome editing tools offers a unique opportunity for targeted therapies. However, effectively normalizing the expre...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Journal Article |
| Published: |
Impact Journals LLC
2016
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| Online Access: | http://hdl.handle.net/20.500.11937/22885 |
| _version_ | 1848750997927624704 |
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| author | Garcia-Bloj, B. Moses, C. Sgro, A. Plani-Lam, J. Arooj, Mahira Duffy, C. Thiruvengadam, S. Sorolla, A. Rashwan, R. Mancera, Ricardo Leisewitz, A. Swift-Scanlan, T. Corvalan, A. Blancafort, P. |
| author_facet | Garcia-Bloj, B. Moses, C. Sgro, A. Plani-Lam, J. Arooj, Mahira Duffy, C. Thiruvengadam, S. Sorolla, A. Rashwan, R. Mancera, Ricardo Leisewitz, A. Swift-Scanlan, T. Corvalan, A. Blancafort, P. |
| author_sort | Garcia-Bloj, B. |
| building | Curtin Institutional Repository |
| collection | Online Access |
| description | The aberrant epigenetic silencing of tumor suppressor genes (TSGs) plays a major role during carcinogenesis and regaining these dormant functions by engineering of sequence-specific epigenome editing tools offers a unique opportunity for targeted therapies. However, effectively normalizing the expression and regaining tumor suppressive functions of silenced TSGs by artificial transcription factors (ATFs) still remains a major challenge. Herein we describe novel combinatorial strategies for the potent reactivation of two class II TSGs, MASPIN and REPRIMO, in cell lines with varying epigenetic states, using the CRISPR/dCas9 associated system linked to a panel of effector domains (VP64, p300, VPR and SAM complex), as well as with protein-based ATFs, Zinc Fingers and TALEs. We found that co-delivery of multiple effector domains using a combination of CRISPR/dCas9 and TALEs or SAM complex maximized activation in highly methylated promoters. In particular, CRISPR/dCas9 VPR with SAM upregulated MASPIN mRNA (22,145-fold change) in H157 lung cancer cells, with accompanying re-expression of MASPIN protein, which led to a concomitant inhibition of cell proliferation and induction of apoptotic cell death. Consistently, CRISPR/dCas9 VP64 with SAM upregulated REPRIMO (680-fold change), which led to phenotypic reprogramming in AGS gastric cancer cells. Altogether, our results outlined novel sequence-specific, combinatorial epigenome editing approaches to reactivate highly methylated TSGs as a promising therapy for cancer and other diseases. |
| first_indexed | 2025-11-14T07:45:44Z |
| format | Journal Article |
| id | curtin-20.500.11937-22885 |
| institution | Curtin University Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-14T07:45:44Z |
| publishDate | 2016 |
| publisher | Impact Journals LLC |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | curtin-20.500.11937-228852017-09-13T13:57:42Z Waking up dormant tumor suppressor genes with zinc fingers, TALEs and the CRISPR/dCas9 system Garcia-Bloj, B. Moses, C. Sgro, A. Plani-Lam, J. Arooj, Mahira Duffy, C. Thiruvengadam, S. Sorolla, A. Rashwan, R. Mancera, Ricardo Leisewitz, A. Swift-Scanlan, T. Corvalan, A. Blancafort, P. The aberrant epigenetic silencing of tumor suppressor genes (TSGs) plays a major role during carcinogenesis and regaining these dormant functions by engineering of sequence-specific epigenome editing tools offers a unique opportunity for targeted therapies. However, effectively normalizing the expression and regaining tumor suppressive functions of silenced TSGs by artificial transcription factors (ATFs) still remains a major challenge. Herein we describe novel combinatorial strategies for the potent reactivation of two class II TSGs, MASPIN and REPRIMO, in cell lines with varying epigenetic states, using the CRISPR/dCas9 associated system linked to a panel of effector domains (VP64, p300, VPR and SAM complex), as well as with protein-based ATFs, Zinc Fingers and TALEs. We found that co-delivery of multiple effector domains using a combination of CRISPR/dCas9 and TALEs or SAM complex maximized activation in highly methylated promoters. In particular, CRISPR/dCas9 VPR with SAM upregulated MASPIN mRNA (22,145-fold change) in H157 lung cancer cells, with accompanying re-expression of MASPIN protein, which led to a concomitant inhibition of cell proliferation and induction of apoptotic cell death. Consistently, CRISPR/dCas9 VP64 with SAM upregulated REPRIMO (680-fold change), which led to phenotypic reprogramming in AGS gastric cancer cells. Altogether, our results outlined novel sequence-specific, combinatorial epigenome editing approaches to reactivate highly methylated TSGs as a promising therapy for cancer and other diseases. 2016 Journal Article http://hdl.handle.net/20.500.11937/22885 10.18632/oncotarget.11142 Impact Journals LLC fulltext |
| spellingShingle | Garcia-Bloj, B. Moses, C. Sgro, A. Plani-Lam, J. Arooj, Mahira Duffy, C. Thiruvengadam, S. Sorolla, A. Rashwan, R. Mancera, Ricardo Leisewitz, A. Swift-Scanlan, T. Corvalan, A. Blancafort, P. Waking up dormant tumor suppressor genes with zinc fingers, TALEs and the CRISPR/dCas9 system |
| title | Waking up dormant tumor suppressor genes with zinc fingers, TALEs and the CRISPR/dCas9 system |
| title_full | Waking up dormant tumor suppressor genes with zinc fingers, TALEs and the CRISPR/dCas9 system |
| title_fullStr | Waking up dormant tumor suppressor genes with zinc fingers, TALEs and the CRISPR/dCas9 system |
| title_full_unstemmed | Waking up dormant tumor suppressor genes with zinc fingers, TALEs and the CRISPR/dCas9 system |
| title_short | Waking up dormant tumor suppressor genes with zinc fingers, TALEs and the CRISPR/dCas9 system |
| title_sort | waking up dormant tumor suppressor genes with zinc fingers, tales and the crispr/dcas9 system |
| url | http://hdl.handle.net/20.500.11937/22885 |