{"doi":"10.7554/elife.53968","title":"Efficient targeted integration directed by short homology in zebrafish and mammalian cells","abstract":"Efficient precision genome engineering requires high frequency and specificity of integration at the genomic target site. Here, we describe a set of resources to streamline reporter gene knock-ins in zebrafish and demonstrate the broader utility of the method in mammalian cells. Our approach uses short homology of 24-48 bp to drive targeted integration of DNA reporter cassettes by homology-mediated end joining (HMEJ) at high frequency at a double strand break in the targeted gene. Our vector series, pGTag (plasmids for Gene Tagging), contains reporters flanked by a universal CRISPR sgRNA sequence which enables in vivo liberation of the homology arms. We observed high rates of germline transmission (22-100%) for targeted knock-ins at eight zebrafish loci and efficient integration at safe harbor loci in porcine and human cells. Our system provides a straightforward and cost-effective approach for high efficiency gene targeting applications in CRISPR and TALEN compatible systems.","journal":"eLife","year":2020,"id":51115,"datarank":3.7671912509293235,"base_score":5.123963979403259,"endowment":5.123963979403259,"self_citation_contribution":0.7685945969104889,"citation_network_contribution":2.9985966540188347,"self_endowment_contribution":0.7685945969104889,"citer_contribution":2.9985966540188347,"corpus_percentile":null,"corpus_rank":null,"citation_count":167,"citer_count":100,"citers_with_citation_signal":100,"citers_with_endowment":100,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9574,"is_data_producer":true,"deposit_databanks":{"Dryad":["10.5061/dryad.m63xsj3zc"]},"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":254316,"name":"Jordan M. 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