title: Targeted repression of AXIN2 and MYC gene expression using designer TALEs.
abstract: Designer TALEs (dTALEs) are chimeric transcription factors that can be engineered to regulate gene expression in mammalian cells. Whether dTALEs can block gene transcription downstream of signal transduction cascades, however, has yet to be fully explored. Here we tested whether dTALEs can be used to target genes whose expression is controlled by Wnt/β-catenin signaling. TALE DNA binding domains were engineered to recognize sequences adjacent to Wnt responsive enhancer elements (WREs) that control expression of axis inhibition protein 2 (AXIN2) and c-MYC (MYC). These custom DNA binding domains were linked to the mSin3A interaction domain (SID) to generate TALE-SID chimeric repressors. The TALE-SIDs repressed luciferase reporter activity, bound their genomic target sites, and repressed AXIN2 and MYC expression in HEK293 cells. We generated a novel HEK293 cell line to determine whether the TALE-SIDs could function downstream of oncogenic Wnt/β-catenin signaling. Treating these cells with doxycycline and tamoxifen stimulates nuclear accumulation of a stabilized form of β-catenin found in a subset of colorectal cancers. The TALE-SIDs repressed AXIN2 and MYC expression in these cells, which suggests that dTALEs could offer an effective therapeutic strategy for the treatment of colorectal cancer.
authors.affiliation: Department of Biochemistry & Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA.
authors.forename: Sherri A
authors.initials: SA
authors.lastname: Rennoll
authors.affiliation: Department of Biochemistry & Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA.
authors.forename: Samantha A
authors.initials: SA
authors.lastname: Scott
authors.affiliation: Department of Biochemistry & Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA. Electronic address: gsy3@psu.edu.
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Yochum
mesh_terms: D060466:Axin Protein / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D015536:Down-Regulation; D016259:Genes, myc; D057809:HEK293 Cells; D006801:Humans; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics*; D015398:Signal Transduction; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D051153:Wnt Proteins / Q000378:metabolism; D051176:beta Catenin / Q000378:metabolism
chemical_list: C491761:AXIN2 protein, human; D060466:Axin Protein; D016271:Proto-Oncogene Proteins c-myc; D014157:Transcription Factors; D051153:Wnt Proteins; D051176:beta Catenin
keywords: AXIN2; MYC; SID; TALE; Wnt; β-Catenin