Herpes ICP8 protein stimulates homologous recombination in human cells
UMass Chan Affiliations
Department of NeurologyDocument Type
Journal ArticlePublication Date
2018-08-15Keywords
Gene targetingRecombinant proteins
DNA recombination
Homologous recombination
293T cells
DNA repair
DNA replication
DNA annealing
Amino Acids, Peptides, and Proteins
Bacteria
Genetic Phenomena
Genetics and Genomics
Viruses
Metadata
Show full item recordAbstract
Recombineering has transformed functional genomic analysis. Genome modification by recombineering using the phage lambda Red homologous recombination protein Beta in Escherichia coli has approached 100% efficiency. While highly efficient in E. coli, recombineering using the Red Synaptase/Exonuclease pair (SynExo) in other organisms declines in efficiency roughly correlating with phylogenetic distance from E. coli. SynExo recombinases are common to double-stranded DNA viruses infecting a variety of organisms, including humans. Human Herpes virus 1 (HHV1) encodes a SynExo comprised of ICP8 synaptase and UL12 exonuclease. In a previous study, the Herpes SynExo was reconstituted in vitro and shown to catalyze a model recombination reaction. Here we describe stimulation of gene targeting to edit a novel fluorescent protein gene in the human genome using ICP8 and compared its efficiency to that of a "humanized" version of Beta protein from phage lambda. ICP8 significantly enhanced gene targeting rates in HEK 293T cells while Beta was not only unable to catalyze recombineering but inhibited gene targeting using endogenous recombination functions, despite both synaptases being well-expressed and localized to the nucleus. This proof of concept encourages developing species-specific SynExo recombinases for genome engineering.Source
PLoS One. 2018 Aug 15;13(8):e0200955. doi: 10.1371/journal.pone.0200955. eCollection 2018. Link to article on publisher's site
DOI
10.1371/journal.pone.0200955Permanent Link to this Item
http://hdl.handle.net/20.500.14038/40759PubMed ID
30110337Related Resources
Rights
Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.Distribution License
http://creativecommons.org/publicdomain/zero/1.0/ae974a485f413a2113503eed53cd6c53
10.1371/journal.pone.0200955
Scopus Count
Collections
Except where otherwise noted, this item's license is described as Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.