Genetically engineering adenoviral vectors for gene therapy.
Adenoviral (Ad) vectors are commonly used for various gene therapy applications. Significant advances in the genetic engineering of Ad vectors in recent years has highlighted their potential for the treatment of metastatic disease. There are several methods to genetically modify the Ad genome to incorporate retargeting peptides which will redirect the natural tropism of the viruses, including homologous recombination in bacteria or yeast. However, homologous recombination in yeast is highly efficient and can be achieved without the need for extensive cloning strategies. In addition, the method does not rely on the presence of unique restriction sites within the Ad genome and the reagents required for this method are widely available and inexpensive. Large plasmids containing the entire adenoviral genome (~36 kbp) can be modified within Saccharomyces cerevisiae yeast and genomes easily rescued in Escherichia coli hosts for analysis or amplification. A method for two-step homologous recombination in yeast is described in this chapter.
The Jenner Institute, University of Oxford, Oxford, UK.
Lynda
L
Coughlan
D000256:Adenoviridae / Q000235:genetics*; D017503:Adenovirus E3 Proteins / Q000235:genetics; D036022:Capsid Proteins / Q000235:genetics; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics; D018014:Gene Transfer Techniques; D005818:Genetic Engineering / Q000379:methods*; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D057809:HEK293 Cells; D059765:Homologous Recombination; D006801:Humans; D014162:Transfection; D014170:Transformation, Genetic; D014779:Virus Replication
D017503:Adenovirus E3 Proteins; D036022:Capsid Proteins; C018315:hexon capsid protein, Adenovirus