title: Engineering synthetic TALE and CRISPR/Cas9 transcription factors for regulating gene expression.
abstract: Engineered DNA-binding proteins that can be targeted to specific sites in the genome to manipulate gene expression have enabled many advances in biomedical research. This includes generating tools to study fundamental aspects of gene regulation and the development of a new class of gene therapies that alter the expression of endogenous genes. Designed transcription factors have entered clinical trials for the treatment of human diseases and others are in preclinical development. High-throughput and user-friendly platforms for designing synthetic DNA-binding proteins present innovative methods for deciphering cell biology and designing custom synthetic gene circuits. We review two platforms for designing synthetic transcription factors for manipulating gene expression: Transcription activator-like effectors (TALEs) and the RNA-guided clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system. We present an overview of each technology and a guide for designing and assembling custom TALE- and CRISPR/Cas9-based transcription factors. We also discuss characteristics of each platform that are best suited for different applications.
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, United States.
authors.forename: Ami M
authors.initials: AM
authors.lastname: Kabadi
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, NC 27708, United States; Institute for Genome Sciences and Policy, Duke University, Durham, NC 27708, United States; Department of Orthopaedic Surgery, Duke University Medical Center, Durham, NC 27710, United States. Electronic address: charles.gersbach@duke.edu.
authors.forename: Charles A
authors.initials: CA
authors.lastname: Gersbach
mesh_terms: D001483:Base Sequence; D064113:CRISPR-Cas Systems / Q000235:genetics*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D005786:Gene Expression Regulation; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D014157:Transcription Factors
keywords: CRISPR; Cas9; Gene regulation; Gene therapy; Genetic reprogramming; Protein engineering; Synthetic biology; TAL effector; TALE; Transcription factor