Details

Autor(en) / Beteiligte

• Garcia-Bloj, Benjamin
• Moses, Colette
• Sgro, Agustin
• Plani-Lam, Janice
• Arooj, Mahira
• Duffy, Ciara
• Thiruvengadam, Shreyas
• Sorolla, Anabel
• Rashwan, Rabab
• Mancera, Ricardo L
• Leisewitz, Andrea
• Swift-Scanlan, Theresa
• Corvalan, Alejandro H
• Blancafort, Pilar

Titel

Waking up dormant tumor suppressor genes with zinc fingers, TALEs and the CRISPR/dCas9 system

Ist Teil von

• Oncotarget, 2016-09, Vol.7 (37), p.60535-60554

Ort / Verlag

United States: Impact Journals LLC

Erscheinungsjahr

2016

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.