Recombinase-Mediated Cassette Exchange Using Adenoviral Vectors

Methods Mol Biol. 2017:1642:127-150. doi: 10.1007/978-1-4939-7169-5_9.

Abstract

Site-specific recombinases are important tools for the modification of mammalian genomes. In conjunction with viral vectors, they can be utilized to mediate site-specific gene insertions in animals and in cell lines which are difficult to transfect. Here we describe a method for the generation and analysis of an adenovirus vector supporting a recombinase-mediated cassette exchange reaction and discuss the advantages and limitations of this approach.

Keywords: Gene expression; Genome engineering; Genomic target; Site-specific recombinase; TCID50.

MeSH terms

  • Caseins / genetics
  • Caseins / metabolism
  • DNA (Cytosine-5-)-Methyltransferase 1 / genetics
  • DNA (Cytosine-5-)-Methyltransferase 1 / metabolism
  • DNA Nucleotidyltransferases / genetics
  • DNA Nucleotidyltransferases / metabolism
  • Dependovirus / genetics*
  • Dependovirus / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Gene Targeting / methods*
  • Genes, Reporter
  • Genetic Vectors / chemistry
  • Genetic Vectors / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • Homologous Recombination*
  • Humans
  • Integrases / genetics*
  • Integrases / metabolism
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mutagenesis, Insertional / methods*
  • Transfection

Substances

  • Caseins
  • Luminescent Proteins
  • enhanced green fluorescent protein
  • fluorescent protein 583
  • Green Fluorescent Proteins
  • DNA (Cytosine-5-)-Methyltransferase 1
  • DNMT1 protein, human
  • Cre recombinase
  • DNA Nucleotidyltransferases
  • Integrases
  • Site-specific recombinase