ArticleNature protocols2011
Sleeping Beauty transposon mutagenesis in rat spermatogonial stem cells.
Article in Nature protocols, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 31 citations in OpenAlex.
- Construction of an Integration Vector with a Chimeric Signal Peptide for the Expression of Monoclonal Antibodies in Mammalian Cells.Current issues in molecular biology · 2024Article
- Rattus norvegicus Spermatogenesis Colony-Forming Assays.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Genetic association and characterization of FSTL5 in isolated clubfoot.Human molecular genetics · 2021Article
- Neuron-Specific Genome Modification in the Adult Rat Brain Using CRISPR-Cas9 Transgenic Rats.Neuron · 2019Article
- Transposons As Tools for Functional Genomics in Vertebrate Models.Trends in genetics : TIG · 2017Review
- Long Evans rat spermatogonial lines are effective germline vectors for transgenic rat production.Transgenic research · 2017Article
- Isolation and cultivation of naive-like human pluripotent stem cells based on HERVH expression.Nature protocols · 2016Article
- Targeted Germline Modifications in Rats Using CRISPR/Cas9 and Spermatogonial Stem Cells.Cell reports · 2015Article
- Impaired myogenic response and autoregulation of cerebral blood flow is rescued in CYP4A1 transgenic Dahl salt-sensitive rat.American journal of physiology. Regulatory, integrative and comparative physiology · 2015Article
- Germline transgenesis in pigs by cytoplasmic microinjection of Sleeping Beauty transposons.Nature protocols · 2014Article
- Germline transgenesis in rabbits by pronuclear microinjection of Sleeping Beauty transposons.Nature protocols · 2014Article
- Germline transgenesis in rodents by pronuclear microinjection of Sleeping Beauty transposons.Nature protocols · 2014Article
- Recent advances in the development of new transgenic animal technology.Cellular and molecular life sciences : CMLS · 2013Review
- Viral transduction of male germline stem cells results in transgene transmission after germ cell transplantation in pigs.Biology of reproduction · 2013Article
- Cell-intrinsic reprogramming capability: gain or loss of pluripotency in germ cells.Reproductive medicine and biology · 2013Review
- Zinc-finger nuclease mediated disruption of Rag1 in the LEW/Ztm rat.BMC immunology · 2012Article
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 3 countries.
Funding
Abstract
We describe an experimental approach for generating mutant alleles in rat spermatogonial stem cells (SSCs) using Sleeping Beauty (SB) transposon-mediated insertional mutagenesis. The protocol is based on mobilization of mutagenic gene-trap transposons from transfected plasmid vectors into the genomes of cultured stem cells. Cells with transposon insertions in expressed genes are selected on the basis of activation of an antibiotic-resistance gene encoded by the transposon. These gene-trap clones are transplanted into the testes of recipient males (either as monoclonal or polyclonal libraries); crossing of these founders with wild-type females allows the insertions to be passed to F(1) progeny. This simple, economic and user-friendly methodological pipeline enables screens for functional gene annotation in the rat, with applicability in other vertebrate models where germ line-competent stem cells have been established. The complete protocol from transfection of SSCs to the genotyping of heterozygous F(1) offspring that harbor genomic SB gene-trap insertions takes 5-6 months.
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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.