{"doi":"10.1101/2025.07.22.665990","title":"Chemogenetic and optogenetic strategies for spatiotemporal control of split-enzyme-based calcium recording","abstract":"ABSTRACT Methods for monitoring physiological changes in cellular Ca 2+ levels have been in high demand for their utility in monitoring neuronal signaling. Recently, we introduced SCANR (Split-Tobacco Etch Virus (TEV) protease Calcium-regulated Neuron Recorder), which reports on Ca 2+ changes in cells through the binding of calmodulin and M13 to reconstitute an active TEV protease. First-generation SCANR marked all of the Ca 2+ spikes that occur throughout the lifetime of the cell, but it did not have a mechanism for controlling the time window in which recording of physiological changes in Ca 2+ occurred. Here, we explore both chemical and light-based strategies for controlling the time and place in which Ca 2+ recording occurs. We describe the adaptation of six popular chemo- and opto-genetics methods for controlling protein activity and subcellular localization to the SCANR system. We report two successful strategies, one that leverages the LOV-Jα optogenetics system for sterically controlling protein interactions and another that employs chemogenetic manipulation of subcellular protein distribution using the FKBP/FRB rapamycin binding pair.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":571030,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.954,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1476905,"name":"Brianna K. Campmier","orcid":null,"position":1,"is_corresponding":false},{"id":1476906,"name":"Akaash Sharma","orcid":null,"position":2,"is_corresponding":false},{"id":999901,"name":"Kelly N. Eckartt","orcid":"0000-0002-0438-6436","position":3,"is_corresponding":false},{"id":827808,"name":"Scott T. Laughlin","orcid":"0000-0002-9134-1200","position":4,"is_corresponding":false},{"id":1475568,"name":"Yunlei Zhang","orcid":"0000-0003-3536-9142","position":0,"is_corresponding":true}],"reference_count":22,"raw_metadata":null,"created_at":"2026-07-19T02:57:11.713851Z","pmid":"40777439","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}