Transcriptional activation of the MICA gene with an engineered CRISPR-Cas9 system.
Major histocompatibility complex class I polypeptide-related sequence A (MICA) is a prototypical NKG2D ligand. Because immune cells, such as natural killer (NK) cells, recognize virally infected or transformed cells and eliminate them through the interaction between NKG2D receptors on NK cells and NKG2D ligands on pathogenic cells, MICA expression levels are associated with NK cell-mediated immunity. Here, we report that an engineered clustered regularly interspaced short palindromic repeats-Cas9-related complex targeting MICA gene promoter sequences activates transcription of the MICA gene from its endogenous locus. Inhibiting microRNA function, which targets the 3' untranslated region of the MICA gene, enhances this activation. These results demonstrate that the combination of Cas9-based transcriptional activators and simultaneous modulation of microRNA function may be a powerful tool for enhancing MICA protein expression and efficient anti-pathogenic cell immunity.
Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Kazuma
K
Sekiba
Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Mari
M
Yamagami
Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan. Electronic address: otsukamo-tky@umin.ac.jp.
Motoyuki
M
Otsuka
Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Tatsunori
T
Suzuki
Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Takahiro
T
Kishikawa
Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Rei
R
Ishibashi
Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Motoko
M
Ohno
Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Masaya
M
Sato
Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Kazuhiko
K
Koike
D020413:3' Untranslated Regions; D001483:Base Sequence; D064113:CRISPR-Cas Systems; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D005818:Genetic Engineering; D056945:Hep G2 Cells; D015395:Histocompatibility Antigens Class I / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007113:Immunity, Innate; D035683:MicroRNAs / Q000235:genetics* / Q000276:immunology; D011401:Promoter Regions, Genetic; D015533:Transcriptional Activation
D020413:3' Untranslated Regions; D015395:Histocompatibility Antigens Class I; C099187:MHC class I-related chain A; D035683:MicroRNAs
CRISPR; MICA; Transcriptional activation; microRNA