{"doi":"10.1101/2021.05.09.443330","title":"Rapid generation of conditional knockout mice using the CRISPR-CAS9 system and electroporation for neuroscience research","abstract":"Abstract The Cre/Loxp-based conditional knockout technology is a powerful tool for gene function analyses by allowing region-time-specific gene manipulation. However, inserting a pair of LoxP cassettes for generating conditional knock-out can be technically challenging and thus time- and resource-consuming. This study proposes an efficient, low-cost method to generate floxed mice using the in vitro fertilization and the CRISPR-Cas9 system over two consecutive generations. This method allowed us to produce floxed mice targeting exon 5 to exon 6 of CaMK1 in a short period, 125 days, using only 16 mice. The efficiency of generating floxed mice was 10%, significantly higher than the conventional ES cell-based method. We directly edited the genome of C57BL/6N fertilized eggs, our target genetic background, to eliminate additional backcrossing steps. We confirmed that the genome of this floxed mouse is responsive to Cre protein. This low-cost, highly efficient method for generating conditional knock-out will facilitate comprehensive, tissue-specific genome analyses.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2021,"id":226644,"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":0.9476,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":718853,"name":"Yuki Hayano","orcid":"0000-0003-0658-023X","position":1,"is_corresponding":false},{"id":657247,"name":"Yoshihisa Nakahata","orcid":"0000-0003-3403-4527","position":2,"is_corresponding":false},{"id":424655,"name":"Yasuhito Ishigaki","orcid":"0000-0003-1012-8430","position":3,"is_corresponding":false},{"id":360815,"name":"Ryohei Yasuda","orcid":"0000-0001-6263-9297","position":4,"is_corresponding":false},{"id":360814,"name":"Hirofumi Nishizono","orcid":"0000-0002-9394-7089","position":0,"is_corresponding":true}],"reference_count":16,"raw_metadata":null,"created_at":"2026-07-18T23:54:38.004707Z","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":[]}