ArticleNucleic acids research2011
Targeted isolation of cloned genomic regions by recombineering for haplotype phasing and isogenic targeting.
Article in Nucleic acids research, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 15 citations in OpenAlex.
- ExoCET: exonuclease in vitro assembly combined with RecET recombination for highly efficient direct DNA cloning from complex genomes.Nucleic acids research · 2018Article
- Revealing complete complex KIR haplotypes phased by long-read sequencing technology.Genes and immunity · 2017Article
- RecET direct cloning and Redαβ recombineering of biosynthetic gene clusters, large operons or single genes for heterologous expression.Nature protocols · 2016Article
- Signatures of Crested Ibis MHC Revealed by Recombination Screening and Short-Reads Assembly Strategy.PloS one · 2016Article
- Haplotype-resolved genome sequencing: experimental methods and applications.Nature reviews. Genetics · 2015Review
- Lambda red mediated gap repair utilizes a novel replicative intermediate in Escherichia coli.PloS one · 2015Article
- Direct cloning of isogenic murine DNA in yeast and relevance of isogenicity for targeting in embryonic stem cells.PloS one · 2013Article
- Determination of haplotypes at structurally complex regions using emulsion haplotype fusion PCR.BMC genomics · 2012Article
- A genome-scale resource for in vivo tag-based protein function exploration in C. elegans.Cell · 2012Article
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Authors and funding
9 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Studying genetic variations in the human genome is important for understanding phenotypes and complex traits, including rare personal variations and their associations with disease. The interpretation of polymorphisms requires reliable methods to isolate natural genetic variations, including combinations of variations, in a format suitable for downstream analysis. Here, we describe a strategy for targeted isolation of large regions (∼35 kb) from human genomes that is also applicable to any genome of interest. The method relies on recombineering to fish out target fosmid clones from pools and thereby circumvents the laborious need to plate and screen thousands of individual clones. To optimize the method, a new highly recombineering-efficient bacterial host, including inducible TrfA for fosmid copy number amplification, was developed. Various regions were isolated from human embryonic stem cell lines and a personal genome, including highly repetitive and duplicated ones. The maternal and paternal alleles at the MECP2/IRAK 1 loci were distinguished based on identification of novel allele-specific single-nucleotide polymorphisms in regulatory regions. Additionally, we applied further recombineering to construct isogenic targeting vectors for patient-specific applications. These methods will facilitate work to understand the linkage between personal variations and disease propensity, as well as possibilities for personal genome surgery.
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Registered trials
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