title: Engineered TAL Effector modulators for the large-scale gain-of-function screening.
abstract: Recent effective use of TAL Effectors (TALEs) has provided an important approach to the design and synthesis of sequence-specific DNA-binding proteins. However, it is still a challenging task to design and manufacture effective TALE modulators because of the limited knowledge of TALE-DNA interactions. Here we synthesized more than 200 TALE modulators and identified two determining factors of transcription activity in vivo: chromatin accessibility and the distance from the transcription start site. The implementation of these modulators in a gain-of-function screen was successfully demonstrated for four cell lines in migration/invasion assays and thus has broad relevance in this field. Furthermore, a novel TALE-TALE modulator was developed to transcriptionally inhibit target genes. Together, these findings underscore the huge potential of these TALE modulators in the study of gene function, reprogramming of cellular behaviors, and even clinical investigation.
authors.affiliation: Biomedical Engineering Department, College of Engineering, Peking University Yan Nan Yuan 60, Beijing 100871, China.
authors.forename: Hanshuo
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Beijing ViewSolid Biotechnology, Beijing 100034, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Li
authors.affiliation: Center for Epigenetics and Chromatin, School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Sha
authors.initials: S
authors.lastname: Hou
authors.affiliation: Biomedical Engineering Department, College of Engineering, Peking University Yan Nan Yuan 60, Beijing 100871, China.
authors.forename: Gancheng
authors.initials: G
authors.lastname: Wang
authors.affiliation: Biomedical Engineering Department, College of Engineering, Peking University Yan Nan Yuan 60, Beijing 100871, China.
authors.forename: Mingjun
authors.initials: M
authors.lastname: Jiang
authors.affiliation: Biomedical Engineering Department, College of Engineering, Peking University Yan Nan Yuan 60, Beijing 100871, China.
authors.forename: Changhong
authors.initials: C
authors.lastname: Sun
authors.affiliation: Beijing ViewSolid Biotechnology, Beijing 100034, China.
authors.forename: Xiongbing
authors.initials: X
authors.lastname: Hu
authors.affiliation: Beijing ViewSolid Biotechnology, Beijing 100034, China.
authors.forename: Fengfeng
authors.initials: F
authors.lastname: Zhuang
authors.affiliation: Biomedical Engineering Department, College of Engineering, Peking University Yan Nan Yuan 60, Beijing 100871, China.
authors.forename: Zhifei
authors.initials: Z
authors.lastname: Dai
authors.affiliation: Center for Epigenetics and Chromatin, School of Life Sciences, Tsinghua University, Beijing 100084, China jzxi@pku.edu.cn jbdai@biomed.tsinghua.edu.cn.
authors.forename: Junbiao
authors.initials: J
authors.lastname: Dai
authors.affiliation: Biomedical Engineering Department, College of Engineering, Peking University Yan Nan Yuan 60, Beijing 100871, China State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Molecular Medicine, Peking University, Beijing 100871, China jzxi@pku.edu.cn jbdai@biomed.tsinghua.edu.cn.
authors.forename: Jianzhong Jeff
authors.initials: JJ
authors.lastname: Xi
mesh_terms: D002460:Cell Line; D002465:Cell Movement; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D006367:HeLa Cells; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics; D006801:Humans; D010770:Phosphotransferases / Q000235:genetics; D015202:Protein Engineering; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D004268:DNA-Binding Proteins; D017405:Herpes Simplex Virus Protein Vmw65; D012097:Repressor Proteins; D014157:Transcription Factors; D010770:Phosphotransferases