A Powerful CRISPR/Cas9‐Based Method for Targeted Transcriptional Activation. Issue 22 (15th April 2016)

• Record Type: Journal Article
• Authors: Katayama, Shota; Moriguchi, Tetsuo; Ohtsu, Naoki; Kondo, Toru
• Is Part Of: Angewandte Chemie international edition. Volume 55:Issue 22(2016)
• Page Start: 6452
• Page End: 6456
• Publication Date: 2016-04-15
• Subjects: CRISPR/Cas9 -- epigenetics -- gene expression -- gene technology -- synthetic biology; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201601708
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: Targeted transcriptional activation of endogenous genes is important for understanding physiological transcriptional networks, synthesizing genetic circuits, and inducing cellular phenotype changes. The CRISPR/Cas9 system has great potential to achieve this purpose, however, it has not yet been successfully used to efficiently activate endogenous genes and induce changes in cellular phenotype. A powerful method for transcriptional activation by using CRISPR/Cas9 was developed. Replacement of a methylated promoter with an unmethylated one by CRISPR/Cas9 was sufficient to activate the expression of the neural cell gene OLIG2 and the embryonic stem cell gene NANOG in HEK293T cells. Moreover, CRISPR/Cas9‐based OLIG2 activation induced the embryonic carcinoma cell line NTERA‐2 to express the neuronal marker βIII‐tubulin. Abstract : Epigenome editing : A novel CRISPR/Cas9‐based method for transcriptional activation through microhomology‐mediated end‐joining (MMEJ) was developed. CRISPR/Cas9 was used to specifically replace the methylated promotor region of a target gene with an unmethylated copy, thereby leading to efficient gene activation at levels sufficient to bring about changes in cell fate.