{"doi":"10.1016/j.xpro.2025.103694","title":"Protocol for cell-type-specific single-cell labeling and manipulation in Drosophila using a sparse driver system","abstract":"Here, we present a protocol for cell-type-specific single-cell labeling and manipulation in Drosophila using a sparse driver system. We describe steps for generating constructs and fly lines, titrating heat-shocked durations for precise temporal control and desired sparsity, and co-expressing multiple transgenes for experiments. We demonstrate that this generalizable toolkit enables tunable sparsity, multi-color staining, single-cell trans-synaptic tracing, single-cell manipulation, and cell-autonomous gene function analysis. For complete details on the use and execution of this protocol, please refer to Xu et al. 1 • Instructions for generating sparse driver constructs and fly lines • Steps to titrate heat-shocked durations for precise temporal control and desired sparsity • Co-express multiple effectors/reporters for morphology, tracing, and gene manipulation Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we present a protocol for cell-type-specific single-cell labeling and manipulation in Drosophila using a sparse driver system. We describe steps for generating constructs and fly lines, titrating heat-shocked durations for precise temporal control and desired sparsity, and co-expressing multiple transgenes for experiments. We demonstrate that this generalizable toolkit enables tunable sparsity, multi-color staining, single-cell trans-synaptic tracing, single-cell manipulation, and cell-autonomous gene function analysis.","journal":"STAR Protocols","year":2025,"id":550297,"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":2,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9547,"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":645060,"name":"Zhuoran Li","orcid":"0009-0007-2979-5678","position":1,"is_corresponding":false},{"id":242246,"name":"Liqun Luo","orcid":"0000-0001-5467-9264","position":2,"is_corresponding":false},{"id":242237,"name":"Chuanyun Xu","orcid":"0009-0002-1138-871X","position":0,"is_corresponding":true}],"reference_count":23,"raw_metadata":null,"created_at":"2026-07-19T02:54:16.596730Z","pmid":"40073018","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":[]}