Impact of essential genes on the success of genome editing experiments generating 3313 new genetically engineered mouse lines

The International Mouse Phenotyping Consortium (IMPC) systematically produces and phenotypes mouse lines with presumptive null mutations to provide insight into gene function. The IMPC now uses the programmable RNA-guided nuclease Cas9 for its increased capacity and flexibility to efficiently generate null alleles in the C57BL/6N strain. In addition to being a valuable novel and accessible research resource, the production of 3313 knockout mouse lines using comparable protocols provides a rich dataset to analyze experimental and biological variables affecting in vivo gene engineering with Cas9. Mouse line production has two critical steps - generation of founders with the desired allele and germline transmission (GLT) of that allele from founders to offspring. A systematic evaluation of the variables impacting success rates identified gene essentiality as the primary factor influencing successful production of null alleles. Collectively, our findings provide best practice recommendations for using Cas9 to generate alleles in mouse essential genes, many of which are orthologs of genes linked to human disease.

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Publication Details

Journal
Scientific Reports
Published
2024-09-30
DOI
https://doi.org/10.1038/s41598-024-72418-8
Citations
6
Primary Topic
CRISPR and Genetic Engineering
Type
article
Field-Weighted Citation Impact
1.40

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article

Impact of essential genes on the success of genome editing experiments generating 3313 new genetically engineered mouse lines

Kathrin Jäger, Diane Gleeson, David Melvin, Guillaume Pavlovic et al.
6 citations
Scientific Reports
CRISPR and Genetic Engineering
1.40
article

Impact of essential genes on the success of genome editing experiments generating 3313 new genetically engineered mouse lines

Kathrin Jäger, Diane Gleeson, David Melvin, Guillaume Pavlovic, Patricia Castellanos-Penton, Amie Creighton, Adam Caulder, Kiran Rajaya, Antonella Galli, Lauri G. Lintott, Skevoulla Christou, Peter D. Price, Charis Cardeno, Lauryl M. J. Nutter, Christopher J. Lelliott, Angelina Gaspero, Harun Kundi, Helen E. Parkinson, Gillian Sleep, Terrence F. Meehan, Peter M. Kutny, Ellen Brown, Ramiro Ramírez‐Solis, Jana Urbanová, Jason D. Heaney, Timothy A. Bell, Greg Clark, Leslie O. Goodwin, Catherine Ingle, Elif F. Acar, Rachel Fell, Stephen A. Murray, Shalini Kamu Reddy, Daniel M. Barrett, Feng He, Hillary Elrick, Sandra Tondat, Jonathan Burvill, Alasdair J. Allan, Charlotte Davis, Susan Allen, Sarah Atkins, Tania Sorg, Edward J. Ryder, Jana Kopkanova, Wendy Gardiner, Rachel Urban, Martin Fray, Romain Lorentz, Radislav Sedláček, Jacqueline K. White, Gemma Codner, Mary E. Dickinson, Mark Griffiths, Fran J. Pike, Elke Malzer, Dan Schiller, Marie‐Christine Birling, Mark Ruhe, Robert E. Braun, Barry P. Rosen, Maribelle Cruz, Graham Duddy, K. C. Kent Lloyd, Matthew Mackenzie, Joanne Doran, Alex Fower, Hannah Wardle‐Jones, Luís Santos, Petr Kašpárek, Nicola Griggs, Vivek Iyer, Asif Nakhuda, Isabel Lorenzo, Francesco J. DeMayo, Michael Woods, Qing Fan-Lan, Denise G. Lanza, Whitney Rich, Valérie Erbs, Kristin Grimsrud, John R. Seavitt, Valerie Laurin, Colin McKerlie, Holly Swash, Lauren Weavers, Evelyn Grau, Yann Hérault, Mohammed Selloum, Mark Thomas, Ryea Maswood, Allan Bradley, David Gannon, Alison Murphy, R. Carter, D. Smedley, Debarati Sethi, Ashley Pawelka, Laurence Schaeffer, Becky Starbuck, Ann‐Marie Mallon, Shaheen Akhtar, Joanna Bottomley, D. E. Archer, Caroline Sinclair, Katharina Boroviak, Alix Schwiening, Steve D. M. Brown, Michelle Stewart, David J. Adams, Kevin A. Peterson, Philippe André, William C. Skarnes, Joshua A. Wood, Joanna Joeng, Janet Kenyon, Marina Gertsenstein, Jorik Loeffler, James Cleak, Susan Kales, Lydia Teboul, Radka Platte, Ozge Danisment, Sara Wells, Peter Matthews, Robin Crawford, Melissa L. Berry, James Bussell, Parjit Kaur, Victoria Ross, Elodie Sins, Marie Hutchison, Shannon Clarke, Abdel Ayadi, Matthew McKay, Christopher V. McCabe, Brandon Willis, Amit Patel, Brendan Doe, Ruth Avery, Christelle Kujath, Jean-Victor Fougerolle, Sandrine Lejeay, Ilya Paulavets, Jan R. Sidiangco, Susan Varley, Tanya Beyetinova, Tim Brendler-Spaeth, Gemma Frake, Chris Godbehere, Sarah Hazeltine
article en
6 citations

Abstract

The International Mouse Phenotyping Consortium (IMPC) systematically produces and phenotypes mouse lines with presumptive null mutations to provide insight into gene function. The IMPC now uses the programmable RNA-guided nuclease Cas9 for its increased capacity and flexibility to efficiently generate null alleles in the C57BL/6N strain. In addition to being a valuable novel and accessible research resource, the production of 3313 knockout mouse lines using comparable protocols provides a rich dataset to analyze experimental and biological variables affecting in vivo gene engineering with Cas9. Mouse line production has two critical steps - generation of founders with the desired allele and germline transmission (GLT) of that allele from founders to offspring. A systematic evaluation of the variables impacting success rates identified gene essentiality as the primary factor influencing successful production of null alleles. Collectively, our findings provide best practice recommendations for using Cas9 to generate alleles in mouse essential genes, many of which are orthologs of genes linked to human disease.

Scientific ReportsVol. 14(1)
Centre National de la Recherche Scientifique (FR), European Bioinformatics Institute (GB), Research Triangle Park Foundation (US), Inserm (FR), Fordham University (US), The University of Texas at San Antonio Health Science Center (US), Queen Mary University of London (GB), University of California, San Francisco (US), Baylor College of Medicine (US), Wellcome/MRC Cambridge Stem Cell Institute (GB), The Francis Crick Institute (GB), Hospital for Sick Children (CA), Wellcome Sanger Institute (GB), University of Oxford (GB), Mary Lyon Centre at MRC Harwell (GB), National Institute of Environmental Health Sciences (US), Jackson Laboratory (US), Phénotypage et ingénierie précliniques des systèmes d’organismes modèles (FR), Czech Academy of Sciences, Institute of Molecular Genetics (CZ), SickKids Foundation (CA), Toronto Centre for Phenogenomics (CA), William Harvey Research Institute (GB), Baylor Genetics (US), University of Manitoba (CA), University of Guelph (CA), University of California, Davis (US), The University of Texas Health Science Center at Houston (US)
Univerzita Karlova v Praze, Genome Canada, Wellcome Trust, European Molecular Biology Laboratory, Ontario Genomics, Agence Nationale de la Recherche, Institut National de la Santé et de la Recherche Médicale, Ministerstvo Školství, Mládeže a Tělovýchovy, Université de Strasbourg, Akademie Věd České Republiky, Centre National de la Recherche Scientifique, PHENOMIN, National Institutes of Health, Medical Research Council, European Regional Development Fund, Czech Centre for Phenogenomics, Institute of Molecular Genetics of the Czech Academy of Sciences, NIH Office of the Director, Common Fund
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