Scientific Reports (Sep 2024)
Impact of essential genes on the success of genome editing experiments generating 3313 new genetically engineered mouse lines
- Hillary Elrick,
- Kevin A. Peterson,
- Brandon J. Willis,
- Denise G. Lanza,
- Elif F. Acar,
- Edward J. Ryder,
- Lydia Teboul,
- Petr Kasparek,
- Marie-Christine Birling,
- David J. Adams,
- Allan Bradley,
- Robert E. Braun,
- Steve D. Brown,
- Adam Caulder,
- Gemma F. Codner,
- Francesco J. DeMayo,
- Mary E. Dickinson,
- Brendan Doe,
- Graham Duddy,
- Marina Gertsenstein,
- Leslie O. Goodwin,
- Yann Hérault,
- Lauri G. Lintott,
- K. C. Kent Lloyd,
- Isabel Lorenzo,
- Matthew Mackenzie,
- Ann-Marie Mallon,
- Colin McKerlie,
- Helen Parkinson,
- Ramiro Ramirez-Solis,
- John R. Seavitt,
- Radislav Sedlacek,
- William C. Skarnes,
- Damien Smedley,
- Sara Wells,
- Jacqueline K. White,
- Joshua A. Wood,
- International Mouse Phenotyping Consortium,
- Stephen A. Murray,
- Jason D. Heaney,
- Lauryl M. J. Nutter
Affiliations
- Hillary Elrick
- The Centre for Phenogenomics
- Kevin A. Peterson
- The Jackson Laboratory
- Brandon J. Willis
- Mouse Biology Program, University of California-Davis
- Denise G. Lanza
- Department of Molecular and Human Genetic, Baylor College of Medicine
- Elif F. Acar
- The Centre for Phenogenomics
- Edward J. Ryder
- Wellcome Sanger Institute
- Lydia Teboul
- The Mary Lyon Centre, MRC Harwell Institute
- Petr Kasparek
- Czech Centre for Phenogenomics, Institute of Molecular Genetics of the Czech Academy of Sciences
- Marie-Christine Birling
- CNRS, INSERM, CELPHEDIA, PHENOMIN, Institut Clinique de la Souris, Université de Strasbourg
- David J. Adams
- Wellcome Sanger Institute
- Allan Bradley
- Wellcome Sanger Institute
- Robert E. Braun
- The Jackson Laboratory
- Steve D. Brown
- MRC Harwell Institute
- Adam Caulder
- The Mary Lyon Centre, MRC Harwell Institute
- Gemma F. Codner
- The Mary Lyon Centre, MRC Harwell Institute
- Francesco J. DeMayo
- Department of Molecular and Cellular Biology, Baylor College of Medicine
- Mary E. Dickinson
- Department of Integrative Physiology, Baylor College of Medicine
- Brendan Doe
- Wellcome Sanger Institute
- Graham Duddy
- Wellcome Sanger Institute
- Marina Gertsenstein
- The Centre for Phenogenomics
- Leslie O. Goodwin
- The Jackson Laboratory
- Yann Hérault
- CNRS, INSERM, CELPHEDIA, PHENOMIN, Institut Clinique de la Souris, Université de Strasbourg
- Lauri G. Lintott
- The Centre for Phenogenomics
- K. C. Kent Lloyd
- Mouse Biology Program, University of California-Davis
- Isabel Lorenzo
- Department of Molecular and Human Genetic, Baylor College of Medicine
- Matthew Mackenzie
- The Mary Lyon Centre, MRC Harwell Institute
- Ann-Marie Mallon
- MRC Harwell Institute
- Colin McKerlie
- The Hospital for Sick Children
- Helen Parkinson
- European Molecular Biology Laboratory-European Bioinformatics Institute
- Ramiro Ramirez-Solis
- Wellcome Sanger Institute
- John R. Seavitt
- The Jackson Laboratory
- Radislav Sedlacek
- Czech Centre for Phenogenomics, Institute of Molecular Genetics of the Czech Academy of Sciences
- William C. Skarnes
- Wellcome Sanger Institute
- Damien Smedley
- William Harvey Research Institute, Queen Mary University of London
- Sara Wells
- The Mary Lyon Centre, MRC Harwell Institute
- Jacqueline K. White
- The Jackson Laboratory
- Joshua A. Wood
- The Jackson Laboratory
- International Mouse Phenotyping Consortium
- Stephen A. Murray
- The Jackson Laboratory
- Jason D. Heaney
- Department of Molecular and Human Genetic, Baylor College of Medicine
- Lauryl M. J. Nutter
- The Centre for Phenogenomics
- DOI
- https://doi.org/10.1038/s41598-024-72418-8
- Journal volume & issue
-
Vol. 14,
no. 1
pp. 1 – 15
Abstract
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.
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