ArticleNature protocols2013
Generation of transgenic mice with megabase-sized human yeast artificial chromosomes by yeast spheroplast-embryonic stem cell fusion.
Article in Nature protocols, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 41 citations in OpenAlex.
- Yeast nuclei-mediated precise delivery of synthetic megabase-scale human DNA into mammalian embryos.Nature protocols · 2026Review
- Synthetic rewriting technologies in mammalian cells.Nature communications · 2026Review
- De novo assembly and delivery of synthetic megabase-scale human DNA into mouse early embryos.Nature methods · 2025Article
- Rapid human genomic DNA cloning into mouse artificial chromosome via direct chromosome transfer from human iPSC and CRISPR/Cas9-mediated translocation.Nucleic acids research · 2024Article
- A supernumerary synthetic chromosome in Komagataella phaffii as a repository for extraneous genetic material.Microbial cell factories · 2023Article
- YLC-assembly: large DNA assembly via yeast life cycle.Nucleic acids research · 2023Article
- Generation of TGFβR2(-1) neoantigen-specific HLA-DR4-restricted T cell receptors for cancer therapy.Journal for immunotherapy of cancer · 2023Article
- The TelN/tos-assisted precise targeting of chromosome segments (TAPE).Journal of advanced research · 2022Article
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- Identification and ranking of recurrent neo-epitopes in cancer.BMC medical genomics · 2019Article
- Genome Writing: Current Progress and Related Applications.Genomics, proteomics & bioinformatics · 2018Review
- Beyond editing to writing large genomes.Nature reviews. Genetics · 2017Article
- Efficient size-independent chromosome delivery from yeast to cultured cell lines.Nucleic acids research · 2017Article
- Synthetic Biology in Cell and Organ Transplantation.Cold Spring Harbor perspectives in biology · 2017Review
- Hydrogel-derived non-precious electrocatalysts for efficient oxygen reduction.Scientific reports · 2015Article
- Generation of a conditionally self-eliminating HAC gene delivery vector through incorporation of a tTAVP64 expression cassette.Nucleic acids research · 2015Article
- Design of a functional cyclic HSV1-TK reporter and its application to PET imaging of apoptosis.Nature protocols · 2015Article
- Targeting cancer-specific mutations by T cell receptor gene therapy.Current opinion in immunology · 2015Review
- Transferring whole genomes from bacteria to yeast spheroplasts using entire bacterial cells to reduce DNA shearing.Nature protocols · 2014Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Introducing human genes into mice offers the opportunity to analyze their in vivo function or to obtain therapeutic molecules. For proper gene regulation, or in case of multigene families, megabase (Mb)-sized DNA fragments often have to be used. Yeast artificial chromosome (YAC)-mediated transgenesis is irreplaceable for this purpose, because alternative methods such as the use of bacterial artificial chromosomes (BACs) cannot introduce DNA fragments larger than 500 kb into the mouse germ line. However, YAC libraries often contain only partial gene loci. Time-consuming reconstruction of YACs, genetic instability and the difficulty in obtaining intact YAC DNA above a certain size impede the generation of humanized mice. Here we describe how to reconstruct YACs containing Mb-sized human DNA, such as the T cell receptor-α (TRA) gene locus, thus facilitating the introduction of large DNA fragments into the mouse germ line. Fusion of YAC-containing yeast and embryonic stem (ES) cells avoids the need for YAC DNA purification. These ES cells are then used to stably introduce the functional TRA gene locus into the mouse germ line. The protocol takes ∼1 year to complete, from reconstruction of the entire TRA gene locus from YACs containing partial but overlapping TRA regions to germline transmission of the YAC.
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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.