title: CRASP: CFP reconstitution across synaptic partners.
abstract: Mapping the pattern of connectivity between neurons is widely regarded to be critical for understanding the nervous system. GRASP (GFP reconstitution across synaptic partners) has been used as a promising method for mapping neuronal connectivity, but is currently available in the green color only, limiting its potential applications. Here we demonstrate CRASP (CFP reconstitution across synaptic partners), a cyan-colored version of GRASP. We validated the system in HEK 293T cells, and generated transgenic Drosophila lines to show that the system could reliably detect neuronal contacts in the brain. Furthermore, we showed that the CRASP signal could be selectively amplified using standard immunohistochemistry methods. The CRASP system adds to the toolkit available to researchers for mapping neuronal connectivity, and substantially expands the potential application of GRASP-like strategies.
authors.affiliation: Institute of Neuroscience, State Key Laboratory of Neuroscience, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Yiming
authors.initials: Y
authors.lastname: Li
authors.affiliation: Institute of Neuroscience, State Key Laboratory of Neuroscience, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China; State Key Laboratory of Brain and Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Aike
authors.initials: A
authors.lastname: Guo
authors.affiliation: Institute of Neuroscience, State Key Laboratory of Neuroscience, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China. Electronic address: hli@ion.ac.cn.
authors.forename: Hao
authors.initials: H
authors.lastname: Li
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D063132:Connectome / Q000379:methods*; D004330:Drosophila; D049452:Green Fluorescent Proteins; D057809:HEK293 Cells; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D009474:Neurons / Q000166:cytology*; D013569:Synapses / Q000648:ultrastructure*
chemical_list: C485182:Cyan Fluorescent Protein; D049452:Green Fluorescent Proteins
keywords: CFP reconstitution; CRASP; Connectome; Drosophila; GFP reconstitution; GRASP