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A Powerful CRISPR/Cas9‐Based Method for Targeted Transcriptional Activation
Author(s) -
Katayama Shota,
Moriguchi Tetsuo,
Ohtsu Naoki,
Kondo Toru
Publication year - 2016
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201601708
Subject(s) - crispr , cas9 , genome editing , hek 293 cells , biology , gene , embryonic stem cell , phenotype , crispr interference , microbiology and biotechnology , homeobox protein nanog , genetics , induced pluripotent stem cell
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.