Recombineering: A Homologous Recombination-Based Method of Genetic Engineering

By S. K. Sharan, L. Thomason, S. Kuznetsov, D. Court. Nature Protocols, 2009

804 citations recorded by the source corpus.

Recombineering is an efficient method of in vivo genetic engineering applicable to chromosomal as well as episomal replicons in Escherichia coli. This method circumvents the need for most standard in vitro cloning techniques. Recombineering allows construction of DNA molecules with precise junctions without constraints being imposed by restriction enzyme site location. Bacteriophage homologous recombination proteins catalyze these recombineering reactions using double- and single-stranded linear DNA substrates, so-called targeting constructs, introduced by electroporation. Gene knockouts, deletions and point mutations are readily made, gene tags can be inserted and regions of bacterial artificial chromosomes or the E. coli genome can be subcloned by gene retrieval using recombineering. Most of these constructs can be made within about 1 week's time.

Subfields: Genetics, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/6793f46f541b6cc775daf5ad6cade41d376d837c

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Citations in the corpus

• Genetic manipulation of genes and cells in the nervous system of the fruit fly (2011, background): “Recombineering can be performed with different recombination templates such as PCR products that encompass protein tags or oligonucleotides that encompass specific mutations (Sharan et al., 2009).”
• Gremlin 1 Identifies a Skeletal Stem Cell with Bone, Cartilage, and Reticular Stromal Potential (2015, methodology): “We generated the Grem1-CreERT transgenic by BAC recombineering (clone RP24-317C19), as previously described (Sharan et al., 2009).”
• Multigene Editing in the Escherichia coli Genome via the CRISPR-Cas9 System (2015, methodology): “pCas were prepared as described previously (16, 36, 37).”