A genome-wide map of adeno-associated virus-mediated human gene targeting

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

To determine which genomic features promote homologous recombination, we created a genome-wide map of gene targeting sites. We used an adeno-associated virus vector to target identical loci introduced as transcriptionally active retroviral vectors. A comparison of ~2,000 targeted and untargeted sites showed that targeting occurred throughout the human genome and was not influenced by the presence of nearby CpG islands, sequence repeats or DNase I-hypersensitive sites. Targeted sites were preferentially located within transcription units, especially when the target loci were transcribed in the opposite orientation to their surrounding chromosomal genes. We determined the impact of DNA replication by mapping replication forks, which revealed a preference for recombination at target loci transcribed toward an incoming fork. Our results constitute the first genome-wide screen of gene targeting in mammalian cells and demonstrate a strong recombinogenic effect of colliding polymerases.

Original languageEnglish (US)
Pages (from-to)969-975
Number of pages7
JournalNature structural & molecular biology
Volume21
Issue number11
DOIs
StatePublished - Nov 1 2014

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Dependovirus
Gene Targeting
Genome
CpG Islands
Deoxyribonuclease I
Homologous Recombination
Human Genome
DNA Replication
Genetic Recombination
Genes

ASJC Scopus subject areas

  • Medicine(all)

Cite this

A genome-wide map of adeno-associated virus-mediated human gene targeting. / Deyle, David R.

In: Nature structural & molecular biology, Vol. 21, No. 11, 01.11.2014, p. 969-975.

Research output: Contribution to journalArticle

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