Document Type
Article
Publication Date
9-30-2024
Original Citation
Elrick H,
Peterson K,
Willis B,
Lanza D,
Acar E,
Ryder E,
Teboul L,
Kasparek P,
Birling M,
Adams D,
Bradley A,
Braun R,
Brown S,
Caulder A,
Codner G,
DeMayo F,
Dickinson M,
Doe B,
Duddy G,
Gertsenstein M,
Goodwin L,
Hérault Y,
Lintott L,
Lloyd K,
Lorenzo I,
Mackenzie M,
Mallon A,
McKerlie C,
Parkinson H,
Ramirez-Solis R,
Seavitt J,
Sedlacek R,
Skarnes W,
Smedley D,
Wells S,
White J,
Wood J,
,
Murray S,
Heaney J,
Nutter L.
Impact of essential genes on the success of genome editing experiments generating 3313 new genetically engineered mouse lines. Sci Rep. 2024;14(1):22626
Keywords
JGM, Animals, Genes, Essential, Mice, Gene Editing, Mice, Knockout, CRISPR-Cas Systems, Alleles, Mice, Inbred C57BL, Male, Female, Genetic Engineering, Phenotype
JAX Source
Sci Rep. 2024;14(1):22626
ISSN
2045-2322
PMID
39349521
DOI
https://doi.org/10.1038/s41598-024-72418-8
Grant
UM1OD023222 supported L.G., K.P., R.B., J.K.W., and S.A.M
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.
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.