{"doi":"10.1007/978-1-0716-2541-5_7","title":"Tissue-Specific CRISPR-Cas9 Screening in Drosophila","abstract":"<jats:title>Abstract</jats:title><jats:p>Over the last century research in <jats:italic>Drosophila</jats:italic> has resulted in many fundamental contributions to our understanding of the biology of multicellular organisms. Many of these breakthroughs have been based on the identification of novel gene functions in large-scale genetic screens. However, conventional forward-genetic screens have been limited by the random nature of mutagenesis and difficulties in mapping causal mutations, while reverse-genetic RNAi screens suffer from incomplete knockdown of gene expression. Recently developed large-scale CRISPR-Cas9 libraries promise to address these limitations by allowing the induction of targeted mutations in genes with spatial and temporal control. Here, we provide a guide for tissue-specific CRISPR screening in <jats:italic>Drosophila</jats:italic>, including the characterization of Gal4 UAS-Cas9 lines, selection of sgRNA libraries, and various quality control measures. We also discuss confounding factors that can give rise to false-positive and false-negative results in such experiments and suggest strategies on how to detect and avoid them. Conditional CRISPR screening represents an exciting new approach for functional genomics in vivo and is set to further expand our knowledge of the molecular underpinning of development, homeostasis, and disease.</jats:p>","journal":"Methods in Molecular Biology","year":2022,"id":602350,"datarank":0.4335557636844247,"base_score":2.8903717578961645,"endowment":2.8903717578961645,"self_citation_contribution":0.4335557636844247,"citation_network_contribution":0.0,"self_endowment_contribution":0.4335557636844247,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":17,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":570,"name":"Michael Boutros","orcid":"0000-0002-9458-817X","position":1,"is_corresponding":false},{"id":564302,"name":"Fillip Port","orcid":"0000-0002-5157-4835","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Tissue-Specific CRISPR-Cas9 Screening in Drosophila","abstract":"<jats:title>Abstract</jats:title><jats:p>Over the last century research in <jats:italic>Drosophila</jats:italic> has resulted in many fundamental contributions to our understanding of the biology of multicellular organisms. Many of these breakthroughs have been based on the identification of novel gene functions in large-scale genetic screens. However, conventional forward-genetic screens have been limited by the random nature of mutagenesis and difficulties in mapping causal mutations, while reverse-genetic RNAi screens suffer from incomplete knockdown of gene expression. Recently developed large-scale CRISPR-Cas9 libraries promise to address these limitations by allowing the induction of targeted mutations in genes with spatial and temporal control. Here, we provide a guide for tissue-specific CRISPR screening in <jats:italic>Drosophila</jats:italic>, including the characterization of Gal4 UAS-Cas9 lines, selection of sgRNA libraries, and various quality control measures. We also discuss confounding factors that can give rise to false-positive and false-negative results in such experiments and suggest strategies on how to detect and avoid them. Conditional CRISPR screening represents an exciting new approach for functional genomics in vivo and is set to further expand our knowledge of the molecular underpinning of development, homeostasis, and disease.</jats:p>","is_dataset_classified":null,"base_score":2.8903717578961645,"endowment":2.8903717578961645,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"35980577","pmcid":null,"openalex_id":"https://openalex.org/W4292231034","authors":[],"funders":[{"funder_name":"European Commission","grant_id":"810296","title":"Decoding Context-Dependent Genetic Networks in vivo"}],"total_grants":1,"fwci":7.9723,"citation_percentile":0.98205591,"influential_citations":0,"citation_trend":[{"year":2023,"count":9},{"year":2024,"count":4},{"year":2025,"count":3},{"year":2026,"count":1}],"oa_status":"hybrid","license":"cc-by","oa_locations":[{"url":"https://link.springer.com/content/pdf/10.1007/978-1-0716-2541-5_7.pdf","host_type":"book series"},{"url":"https://link.springer.com/content/pdf/10.1007/978-1-0716-2541-5_7.pdf","host_type":"publisher"},{"url":"https://link.springer.com/content/pdf/10.1007/978-1-0716-2541-5_7","host_type":"publisher"},{"url":"https://doi.org/10.1007/978-1-0716-2541-5_7","host_type":"book series"},{"url":"https://pubmed.ncbi.nlm.nih.gov/35980577","host_type":"repository"},{"url":"http://dx.doi.org/10.1007/978-1-0716-2541-5_7","host_type":""}],"fields_of_study":["CRISPR and Genetic Engineering","Genetics, Aging, and Longevity in Model Organisms","Chromosomal and Genetic Variations","0301 basic medicine","0303 health sciences","03 medical and health sciences","Animals","CRISPR-Cas Systems","Drosophila","Genetic Testing","Genomics","Mutagenesis","RNA, Small Untranslated"],"mesh_terms":["Animals","Drosophila","Genetic Testing","Mutagenesis","Genomics","RNA, Small Untranslated","CRISPR-Cas Systems"],"keywords":["CRISPR","Biology","Computational biology","Cas9","Genetic screen","Forward genetics","Multicellular organism","Genetics","Mutagenesis","Gene knockdown","Functional genomics","Gene","RNA interference","Genome editing","Genomics","Mutation","Genome","Phenotype","RNA","Drosophila","Screening","Crispr-cas9","Sgrna Libraries","Animals","RNA, Small Untranslated","Genetic Testing","CRISPR-Cas Systems"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-29T19:08:25.376132Z","pmid":null,"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":[]}