{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T03:29:47Z","timestamp":1777346987760,"version":"3.51.4"},"reference-count":47,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,12,6]],"date-time":"2025-12-06T00:00:00Z","timestamp":1764979200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T00:00:00Z","timestamp":1768435200000},"content-version":"vor","delay-in-days":40,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["220540\/Z\/20\/A"],"award-info":[{"award-number":["220540\/Z\/20\/A"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["220540\/Z\/20\/A"],"award-info":[{"award-number":["220540\/Z\/20\/A"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["220540\/Z\/20\/A"],"award-info":[{"award-number":["220540\/Z\/20\/A"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["220540\/Z\/20\/A"],"award-info":[{"award-number":["220540\/Z\/20\/A"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["220540\/Z\/20\/A"],"award-info":[{"award-number":["220540\/Z\/20\/A"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["220540\/Z\/20\/A"],"award-info":[{"award-number":["220540\/Z\/20\/A"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["220540\/Z\/20\/A"],"award-info":[{"award-number":["220540\/Z\/20\/A"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["220540\/Z\/20\/A"],"award-info":[{"award-number":["220540\/Z\/20\/A"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["220540\/Z\/20\/A"],"award-info":[{"award-number":["220540\/Z\/20\/A"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["220540\/Z\/20\/A"],"award-info":[{"award-number":["220540\/Z\/20\/A"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["220540\/Z\/20\/A"],"award-info":[{"award-number":["220540\/Z\/20\/A"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"name":"OpenTargets"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Nat Commun"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Pairwise perturbation of gene function using the CRISPR\/Cas9 system has potential in screening for genetic interactions and synthetic lethal gene pairs to identify combination therapies for cancer. However, existing dual guide expression systems are cumbersome to clone, often result in a large proportion of undesired guide pairs and have an imbalance of guide expression from the two positions. Here, we demonstrate a next-generation system for dual guide delivery based around a tRNA spacer that allows a single-step cloning strategy, as little as 2% of undesired guide pairs, and highly balanced expression of the two guides. This system allows efficient library-scale screening for hundreds of thousands of genetic interactions using the well-understood\n                    <jats:italic>Streptococcus pyogenes<\/jats:italic>\n                    Cas9 (SpCas9) system. We use this to screen a 100,136 guide pair library in colorectal cancer cells and successfully identify synthetic lethal genetic interactions between paralogs or other known interacting genes, establishing our method for performing efficient large-scale genetic interaction screens. This system is versatile and could be used with most guide RNA vector systems, and for other uses of paired guide delivery, such as improving single gene knockout efficiency or improving guide detection in single cell or optical CRISPR screens.\n                  <\/jats:p>","DOI":"10.1038\/s41467-025-67256-9","type":"journal-article","created":{"date-parts":[[2025,12,6]],"date-time":"2025-12-06T11:04:13Z","timestamp":1765019053000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["A next-generation dual guide CRISPR system for genetic interaction library screening"],"prefix":"10.1038","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-8324-8934","authenticated-orcid":false,"given":"Thomas","family":"Burgold","sequence":"first","affiliation":[]},{"given":"Emre","family":"Karakoc","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9967-5205","authenticated-orcid":false,"given":"Emanuel","family":"Gon\u00e7alves","sequence":"additional","affiliation":[]},{"given":"Inigo","family":"Barrio-Hernandez","sequence":"additional","affiliation":[]},{"given":"Lisa","family":"Dwane","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5429-0152","authenticated-orcid":false,"given":"Romina","family":"Silva","sequence":"additional","affiliation":[]},{"given":"Emily","family":"Souster","sequence":"additional","affiliation":[]},{"given":"Mamta","family":"Sharma","sequence":"additional","affiliation":[]},{"given":"Alexandra","family":"Beck","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3840-4954","authenticated-orcid":false,"given":"Gene Ching Chiek","family":"Koh","sequence":"additional","affiliation":[]},{"given":"Lykourgos-Panagiotis","family":"Zalmas","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2618-4237","authenticated-orcid":false,"given":"Mathew J.","family":"Garnett","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1632-9137","authenticated-orcid":false,"given":"Andrew R.","family":"Bassett","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2025,12,6]]},"reference":[{"key":"67256_CR1","doi-asserted-by":"publisher","first-page":"511","DOI":"10.1038\/s41586-019-1103-9","volume":"568","author":"FM Behan","year":"2019","unstructured":"Behan, F. M. et al. Prioritization of cancer therapeutic targets using CRISPR-Cas9 screens. Nature 568, 511\u2013516 (2019).","journal-title":"Nature"},{"key":"67256_CR2","doi-asserted-by":"publisher","first-page":"551","DOI":"10.1038\/s41586-019-1102-x","volume":"568","author":"EM Chan","year":"2019","unstructured":"Chan, E. M. et al. WRN helicase is a synthetic lethal target in microsatellite unstable cancers. Nature 568, 551\u2013556 (2019).","journal-title":"Nature"},{"key":"67256_CR3","doi-asserted-by":"publisher","first-page":"e0187236","DOI":"10.1371\/journal.pone.0187236","volume":"12","author":"F Adikusuma","year":"2017","unstructured":"Adikusuma, F., Pfitzner, C. & Thomas, P. Q. Versatile single-step-assembly CRISPR\/Cas9 vectors for dual gRNA expression. PLoS One 12, e0187236 (2017).","journal-title":"PLoS One"},{"key":"67256_CR4","doi-asserted-by":"publisher","first-page":"573","DOI":"10.1038\/nmeth.4225","volume":"14","author":"JP Shen","year":"2017","unstructured":"Shen, J. P. et al. Combinatorial CRISPR-Cas9 screens for de novo mapping of genetic interactions. Nat. Methods 14, 573\u2013576 (2017).","journal-title":"Nat. Methods"},{"key":"67256_CR5","doi-asserted-by":"publisher","first-page":"348","DOI":"10.1016\/j.molcel.2017.06.030","volume":"67","author":"N Erard","year":"2017","unstructured":"Erard, N., Knott, S. R. V. & Hannon, G. J. A CRISPR Resource for Individual, Combinatorial, or Multiplexed Gene Knockout. Mol. Cell 67, 348\u2013354.e4 (2017).","journal-title":"Mol. Cell"},{"key":"67256_CR6","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms9083","volume":"6","author":"JA Vidigal","year":"2015","unstructured":"Vidigal, J. A. & Ventura, A. Rapid and efficient one-step generation of paired gRNA CRISPR-Cas9 libraries. Nat. Commun. 6, 8083 (2015).","journal-title":"Nat. Commun."},{"key":"67256_CR7","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-21478-9","volume":"12","author":"NA Thompson","year":"2021","unstructured":"Thompson, N. A. et al. Combinatorial CRISPR screen identifies fitness effects of gene paralogues. Nat. Commun. 12, 1302 (2021).","journal-title":"Nat. Commun."},{"key":"67256_CR8","doi-asserted-by":"publisher","first-page":"5684","DOI":"10.1093\/nar\/gkab309","volume":"49","author":"V Diehl","year":"2021","unstructured":"Diehl, V. et al. Minimized combinatorial CRISPR screens identify genetic interactions in autophagy. Nucleic Acids Res 49, 5684\u20135704 (2021).","journal-title":"Nucleic Acids Res"},{"key":"67256_CR9","doi-asserted-by":"publisher","first-page":"271","DOI":"10.1038\/nmeth.4604","volume":"15","author":"AJ Hill","year":"2018","unstructured":"Hill, A. J. et al. On the design of CRISPR-based single-cell molecular screens. Nat. Methods 15, 271\u2013274 (2018).","journal-title":"Nat. Methods"},{"key":"67256_CR10","doi-asserted-by":"publisher","first-page":"e0197547","DOI":"10.1371\/journal.pone.0197547","volume":"13","author":"M Hegde","year":"2018","unstructured":"Hegde, M., Strand, C., Hanna, R. E. & Doench, J. G. Uncoupling of sgRNAs from their associated barcodes during PCR amplification of combinatorial CRISPR screens. PLoS One 13, e0197547 (2018).","journal-title":"PLoS One"},{"key":"67256_CR11","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1038\/nbt.4048","volume":"36","author":"FJ Najm","year":"2018","unstructured":"Najm, F. J. et al. Orthologous CRISPR-Cas9 enzymes for combinatorial genetic screens. Nat. Biotechnol. 36, 179\u2013189 (2018).","journal-title":"Nat. Biotechnol."},{"key":"67256_CR12","doi-asserted-by":"publisher","first-page":"94","DOI":"10.1038\/s41587-020-0600-6","volume":"39","author":"PC DeWeirdt","year":"2021","unstructured":"DeWeirdt, P. C. et al. Optimization of AsCas12a for combinatorial genetic screens in human cells. Nat. Biotechnol. 39, 94\u2013104 (2021).","journal-title":"Nat. Biotechnol."},{"key":"67256_CR13","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-30196-9","volume":"13","author":"R Li","year":"2022","unstructured":"Li, R. et al. Comparative optimization of combinatorial CRISPR screens. Nat. Commun. 13, 2469 (2022).","journal-title":"Nat. Commun."},{"key":"67256_CR14","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41467-020-17209-1","volume":"11","author":"RA Gier","year":"2020","unstructured":"Gier, R. A. et al. High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening. Nat. Commun. 11, 1\u20139 (2020).","journal-title":"Nat. Commun."},{"key":"67256_CR15","doi-asserted-by":"publisher","first-page":"239","DOI":"10.1038\/nbt.4061","volume":"36","author":"HK Kim","year":"2018","unstructured":"Kim, H. K. et al. Deep learning improves prediction of CRISPR-Cpf1 guide RNA activity. Nat. Biotechnol. 36, 239\u2013241 (2018).","journal-title":"Nat. Biotechnol."},{"key":"67256_CR16","doi-asserted-by":"publisher","first-page":"276","DOI":"10.1038\/s41587-018-0011-0","volume":"37","author":"BP Kleinstiver","year":"2019","unstructured":"Kleinstiver, B. P. et al. Engineered CRISPR-Cas12a variants with increased activities and improved targeting ranges for gene, epigenetic and base editing. Nat. Biotechnol. 37, 276\u2013282 (2019).","journal-title":"Nat. Biotechnol."},{"key":"67256_CR17","doi-asserted-by":"publisher","first-page":"759","DOI":"10.1016\/j.cell.2015.09.038","volume":"163","author":"B Zetsche","year":"2015","unstructured":"Zetsche, B. et al. Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell 163, 759\u2013771 (2015).","journal-title":"Cell"},{"key":"67256_CR18","doi-asserted-by":"publisher","first-page":"852","DOI":"10.1038\/nmeth.3972","volume":"13","author":"F Port","year":"2016","unstructured":"Port, F. & Bullock, S. L. Augmenting CRISPR applications in Drosophila with tRNA-flanked sgRNAs. Nat. Methods 13, 852\u2013854 (2016).","journal-title":"Nat. Methods"},{"key":"67256_CR19","doi-asserted-by":"publisher","first-page":"3570","DOI":"10.1073\/pnas.1420294112","volume":"112","author":"K Xie","year":"2015","unstructured":"Xie, K., Minkenberg, B. & Yang, Y. Boosting CRISPR\/Cas9 multiplex editing capability with the endogenous tRNA-processing system. Proc. Natl. Acad. Sci. Usa. 112, 3570\u20133575 (2015).","journal-title":"Proc. Natl. Acad. Sci. Usa."},{"key":"67256_CR20","doi-asserted-by":"publisher","first-page":"889","DOI":"10.1016\/j.bbrc.2016.11.129","volume":"482","author":"F Dong","year":"2017","unstructured":"Dong, F., Xie, K., Chen, Y., Yang, Y. & Mao, Y. Polycistronic tRNA and CRISPR guide-RNA enables highly efficient multiplexed genome engineering in human cells. Biochem. Biophys. Res. Commun. 482, 889\u2013895 (2017).","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"67256_CR21","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-51090-3","volume":"9","author":"Y Zhao","year":"2019","unstructured":"Zhao, Y. et al. A one-step tRNA-CRISPR system for genome-wide genetic interaction mapping in mammalian cells. Sci. Rep. 9, 14499 (2019).","journal-title":"Sci. Rep."},{"key":"67256_CR22","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-09148-3","volume":"10","author":"DJHF Knapp","year":"2019","unstructured":"Knapp, D. J. H. F. et al. Decoupling tRNA promoter and processing activities enables specific Pol-II Cas9 guide RNA expression. Nat. Commun. 10, 1490 (2019).","journal-title":"Nat. Commun."},{"key":"67256_CR23","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-30514-1","volume":"13","author":"Q Yuan","year":"2022","unstructured":"Yuan, Q. & Gao, X. Multiplex base- and prime-editing with drive-and-process CRISPR arrays. Nat. Commun. 13, 2771 (2022).","journal-title":"Nat. Commun."},{"key":"67256_CR24","doi-asserted-by":"publisher","first-page":"1193","DOI":"10.1016\/j.celrep.2016.09.079","volume":"17","author":"K Tzelepis","year":"2016","unstructured":"Tzelepis, K. et al. A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and Therapeutic Targets in Acute Myeloid Leukemia. Cell Rep. 17, 1193\u20131205 (2016).","journal-title":"Cell Rep."},{"key":"67256_CR25","doi-asserted-by":"publisher","first-page":"463","DOI":"10.1038\/nbt.3834","volume":"35","author":"K Han","year":"2017","unstructured":"Han, K. et al. Synergistic drug combinations for cancer identified in a CRISPR screen for pairwise genetic interactions. Nat. Biotechnol. 35, 463 (2017).","journal-title":"Nat. Biotechnol."},{"key":"67256_CR26","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-023-03132-3","volume":"25","author":"S-J Heo","year":"2024","unstructured":"Heo, S.-J. et al. Compact CRISPR genetic screens enabled by improved guide RNA library cloning. Genome Biol. 25, 25 (2024).","journal-title":"Genome Biol."},{"key":"67256_CR27","doi-asserted-by":"publisher","unstructured":"Walton, R. T., Qin, Y. & Blainey, P. C. CROPseq-multi: a versatile solution for multiplexed perturbation and decoding in pooled CRISPR screens. bioRxiv https:\/\/doi.org\/10.1101\/2024.03.17.585235 (2024).","DOI":"10.1101\/2024.03.17.585235"},{"key":"67256_CR28","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-021-02268-4","volume":"22","author":"E Gon\u00e7alves","year":"2021","unstructured":"Gon\u00e7alves, E. et al. Minimal genome-wide human CRISPR-Cas9 library. Genome Biol. 22, 40 (2021).","journal-title":"Genome Biol."},{"key":"67256_CR29","doi-asserted-by":"publisher","first-page":"e0198635","DOI":"10.1371\/journal.pone.0198635","volume":"13","author":"S Xie","year":"2018","unstructured":"Xie, S., Cooley, A., Armendariz, D., Zhou, P. & Hon, G. C. Frequent sgRNA-barcode recombination in single-cell perturbation assays. PLoS One 13, e0198635 (2018).","journal-title":"PLoS One"},{"key":"67256_CR30","doi-asserted-by":"publisher","unstructured":"Feldman, D., Singh, A., Garrity, A. J. & Blainey, P. C. Lentiviral co-packaging mitigates the effects of intermolecular recombination and multiple integrations in pooled genetic screens. bioRxiv 262121 https:\/\/doi.org\/10.1101\/262121 (2018).","DOI":"10.1101\/262121"},{"key":"67256_CR31","doi-asserted-by":"publisher","unstructured":"Adamson, B., Norman, T. M., Jost, M. & Weissman, J. S. Approaches to maximize sgRNA-barcode coupling in Perturb-seq screens. bioRxiv 298349 https:\/\/doi.org\/10.1101\/298349 (2018)","DOI":"10.1101\/298349"},{"key":"67256_CR32","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-019-1745-9","volume":"20","author":"M Zamanighomi","year":"2019","unstructured":"Zamanighomi, M. et al. GEMINI: a variational Bayesian approach to identify genetic interactions from combinatorial CRISPR screens. Genome Biol. 20, 137 (2019).","journal-title":"Genome Biol."},{"key":"67256_CR33","doi-asserted-by":"crossref","unstructured":"Dede, M., McLaughlin, M., Kim, E. & Hart, T. Multiplex enCas12a screens detect functional buffering among paralogs otherwise masked in monogenic Cas9 knockout screens. Genome Biol. 21, (2020).","DOI":"10.1186\/s13059-020-02173-2"},{"key":"67256_CR34","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms15178","volume":"8","author":"DW Morgens","year":"2017","unstructured":"Morgens, D. W. et al. Genome-scale measurement of off-target activity using Cas9 toxicity in high-throughput screens. Nat. Commun. 8, 15178 (2017).","journal-title":"Nat. Commun."},{"key":"67256_CR35","doi-asserted-by":"publisher","unstructured":"Anzalone, A. V. et al. Search-and-replace genome editing without double-strand breaks or donor DNA. Nature https:\/\/doi.org\/10.1038\/s41586-019-1711-4 (2019).","DOI":"10.1038\/s41586-019-1711-4"},{"key":"67256_CR36","doi-asserted-by":"publisher","first-page":"239","DOI":"10.1016\/j.neuron.2019.07.014","volume":"104","author":"R Tian","year":"2019","unstructured":"Tian, R. et al. CRISPR Interference-Based Platform for Multimodal Genetic Screens in Human iPSC-Derived Neurons. Neuron 104, 239\u2013255.e12 (2019).","journal-title":"Neuron"},{"key":"67256_CR37","doi-asserted-by":"publisher","first-page":"954","DOI":"10.1038\/s41587-020-0470-y","volume":"38","author":"JM Replogle","year":"2020","unstructured":"Replogle, J. M. et al. Combinatorial single-cell CRISPR screens by direct guide RNA capture and targeted sequencing. Nat. Biotechnol. 38, 954\u2013961 (2020).","journal-title":"Nat. Biotechnol."},{"key":"67256_CR38","doi-asserted-by":"publisher","first-page":"2503","DOI":"10.1016\/j.cell.2021.03.025","volume":"184","author":"JK Nu\u00f1ez","year":"2021","unstructured":"Nu\u00f1ez, J. K. et al. Genome-wide programmable transcriptional memory by CRISPR-based epigenome editing. Cell 184, 2503\u20132519.e17 (2021).","journal-title":"Cell"},{"key":"67256_CR39","doi-asserted-by":"publisher","unstructured":"Replogle, J. M. et al. Maximizing CRISPRi efficacy and accessibility with dual-sgRNA libraries and optimal effectors. https:\/\/doi.org\/10.7554\/eLife.81856 (2022).","DOI":"10.7554\/eLife.81856"},{"key":"67256_CR40","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1007\/978-1-4939-9170-9_11","volume":"1961","author":"M Bruntraeger","year":"2019","unstructured":"Bruntraeger, M., Byrne, M., Long, K. & Bassett, A. R. Editing the Genome of Human Induced Pluripotent Stem Cells Using CRISPR\/Cas9 Ribonucleoprotein Complexes. Methods Mol. Biol. 1961, 153\u2013183 (2019).","journal-title":"Methods Mol. Biol."},{"key":"67256_CR41","doi-asserted-by":"publisher","first-page":"695","DOI":"10.1038\/nbt.3583","volume":"34","author":"L Pinello","year":"2016","unstructured":"Pinello, L. et al. Analyzing CRISPR genome-editing experiments with CRISPResso. Nat. Biotechnol. 34, 695\u2013697 (2016).","journal-title":"Nat. Biotechnol."},{"key":"67256_CR42","doi-asserted-by":"publisher","first-page":"1096","DOI":"10.1126\/science.aac7041","volume":"350","author":"T Wang","year":"2015","unstructured":"Wang, T. et al. Identification and characterization of essential genes in the human genome. Science 350, 1096\u20131101 (2015).","journal-title":"Science"},{"key":"67256_CR43","doi-asserted-by":"publisher","first-page":"3078","DOI":"10.1093\/bioinformatics\/btv308","volume":"31","author":"A Hodgkins","year":"2015","unstructured":"Hodgkins, A. et al. WGE: a CRISPR database for genome engineering. Bioinformatics 31, 3078\u20133080 (2015).","journal-title":"Bioinformatics"},{"key":"67256_CR44","doi-asserted-by":"publisher","unstructured":"Barrio-Hernandez, I. et al. ibarrioh\/DualGuide_COLO1: Release2-final (Version Release2), Zenodo, https:\/\/doi.org\/10.5281\/zenodo.17191951 (2025).","DOI":"10.5281\/zenodo.17191951"},{"key":"67256_CR45","doi-asserted-by":"publisher","unstructured":"Barrio-Hernandez, I. et al. MAPPING.zip dataset, Figshare, https:\/\/doi.org\/10.6084\/m9.figshare.25533091.v1 (2024).","DOI":"10.6084\/m9.figshare.25533091.v1"},{"key":"67256_CR46","doi-asserted-by":"publisher","first-page":"2281","DOI":"10.1038\/nprot.2013.143","volume":"8","author":"FA Ran","year":"2013","unstructured":"Ran, F. A. et al. Genome engineering using the CRISPR-Cas9 system. Nat. Protoc. 8, 2281\u20132308 (2013).","journal-title":"Nat. Protoc."},{"key":"67256_CR47","doi-asserted-by":"publisher","first-page":"819","DOI":"10.1126\/science.1231143","volume":"339","author":"L Cong","year":"2013","unstructured":"Cong, L. et al. Multiplex genome engineering using CRISPR\/Cas systems. Science 339, 819\u2013823 (2013).","journal-title":"Science"}],"container-title":["Nature Communications"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41467-025-67256-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41467-025-67256-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41467-025-67256-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T02:02:34Z","timestamp":1768528954000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41467-025-67256-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,6]]},"references-count":47,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["67256"],"URL":"https:\/\/doi.org\/10.1038\/s41467-025-67256-9","relation":{},"ISSN":["2041-1723"],"issn-type":[{"value":"2041-1723","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,6]]},"assertion":[{"value":"23 May 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"25 November 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"6 December 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"A.B. is a founder of and consultant for Ensocell therapeutics. M.G. is a founder of and consultant for Mosaic Therapeutics, receives research funding from GSK and Astex Pharmaceuticals and is a consultant for Bristol-Myers Squibb. The remaining authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"561"}}