title: A novel luciferase knock-in reporter system for studying transcriptional regulation of the human Sox2 gene.
abstract: Sox2 is an important transcriptional factor that has multiple functions in stem cell maintenance and tumorigenesis. To investigate the transcriptional regulation of the Sox2 gene, a luciferase knock-in reporter system was established in HEK293 cells by placing the luciferase gene in the genome under the control of the Sox2 gene promoter using a transcription activator-like effector nuclease (TALEN)-mediated genome editing technique. PCR and Southern blot results confirmed the site-specific integration of a single copy of the exogenous luciferase gene into the genome. To prove the reliability and sensitivity of this novel luciferase knock-in system, a CRISPR/Cas transcription activation system for the Sox2 gene was constructed and applied to the knock-in system. The results indicated that luciferase activity was directly correlated with the activity of the Sox2 endogenous promoter. This novel system will be a useful tool to study the transcriptional regulation of Sox2, and has great potential in medical and industrial applications.
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an 710062, Shaanxi, PR China.
authors.forename: Dan
authors.initials: D
authors.lastname: Xiao
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an 710062, Shaanxi, PR China.
authors.forename: Weifeng
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an 710062, Shaanxi, PR China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an 710062, Shaanxi, PR China.
authors.forename: Kuan
authors.initials: K
authors.lastname: Liu
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an 710062, Shaanxi, PR China.
authors.forename: Junli
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an 710062, Shaanxi, PR China.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Sun
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an 710062, Shaanxi, PR China.
authors.forename: Linlin
authors.initials: L
authors.lastname: Shan
authors.affiliation: Department of Pathology, Northwestern University Feinberg School of Medicine, 303 E. Chicago Avenue, Ward 3-140, Chicago, IL 60611, USA.
authors.forename: Qinwen
authors.initials: Q
authors.lastname: Mao
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an 710062, Shaanxi, PR China. Electronic address: hbxia2001@163.com.
authors.forename: Haibin
authors.initials: H
authors.lastname: Xia
mesh_terms: D064113:CRISPR-Cas Systems; D005786:Gene Expression Regulation; D055879:Gene Knock-In Techniques / Q000379:methods*; D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D055748:SOXB1 Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D008156:Luciferases
keywords: Gene editing; Homologous recombination; Knock-in; Sox2; TALEN