ArticleGenetics2013
Highly efficient targeted mutagenesis in mice using TALENs.
Article in Genetics, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed, 72 citations in OpenAlex.
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- Multiple pathways of toxicity induced byeLife · 2021Article
- C9orf72-mediated ALS and FTD: multiple pathways to disease.Nature reviews. Neurology · 2018Review
- Biological Spectrum of Amyotrophic Lateral Sclerosis Prions.Cold Spring Harbor perspectives in medicine · 2017Review
- A history of genome editing in mammals.Mammalian genome : official journal of the International Mammalian Genome Society · 2017Review
- Mouse Genome Informatics (MGI) Resource: Genetic, Genomic, and Biological Knowledgebase for the Laboratory Mouse.ILAR journal · 2017Review
- Next-generation mammalian genetics toward organism-level systems biology.NPJ systems biology and applications · 2017Review
- Generation of knockout mice by Cpf1-mediated gene targeting.Nature biotechnology · 2016Article
- Gene editing for corneal disease management.World journal of translational medicine · 2016Article
- RS-1 enhances CRISPR/Cas9- and TALEN-mediated knock-in efficiency.Nature communications · 2016Article
- Genome Engineering with TALE and CRISPR Systems in Neuroscience.Frontiers in genetics · 2016Review
- C9orf72 ablation in mice does not cause motor neuron degeneration or motor deficits.Annals of neurology · 2015Article
- Development of an intein-mediated split-Cas9 system for gene therapy.Nucleic acids research · 2015Article
Corrections and comments
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Authors and funding
8 authors at 4 institutions in 1 country.
Funding
Abstract
Targeted mouse mutants are instrumental for the analysis of gene function in health and disease. We recently provided proof-of-principle for the fast-track mutagenesis of the mouse genome, using transcription activator-like effector nucleases (TALENs) in one-cell embryos. Here we report a routine procedure for the efficient production of disease-related knockin and knockout mutants, using improved TALEN mRNAs that include a plasmid-coded poly(A) tail (TALEN-95A), circumventing the problematic in vitro polyadenylation step. To knock out the C9orf72 gene as a model of frontotemporal lobar degeneration, TALEN-95A mutagenesis induced sequence deletions in 41% of pups derived from microinjected embryos. Using TALENs together with mutagenic oligodeoxynucleotides, we introduced amyotrophic lateral sclerosis patient-derived missense mutations in the fused in sarcoma (Fus) gene at a rate of 6.8%. For the simple identification of TALEN-induced mutants and their progeny we validate high-resolution melt analysis (HRMA) of PCR products as a sensitive and universal genotyping tool. Furthermore, HRMA of off-target sites in mutant founder mice revealed no evidence for undesired TALEN-mediated processing of related genomic sequences. The combination of TALEN-95A mRNAs for enhanced mutagenesis and of HRMA for simplified genotyping enables the accelerated, routine production of new mouse models for the study of genetic disease mechanisms.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.