Transgenesis With High Capacity Vectors
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Lecture 26 Transgenesis with high capacity vectors
Transgenesis with High-Capacity Vectors • transgenes are complementary DNAs (cDNAs), small genes (<20 kb) or parts of genes
• cDNAs are poorly expressed in mammalian cells • In transgenesis, important gene- specific regulatory sequences retained as part of the insert
• • complete genes and multigene complexes (<100 kb) large for conventional vectors
• need for high-capacity vectors
• eg. bacterial, P1 bacteriophage- derived, mammalian, and yeast artificial chromosomes [ BACs, PACs,MACs and YACs, respectively])
• carry genomic DNA ranging in size from 100 to >1,000kb • used for transgenesis
YAC transgenesis • transgenic mice produced by microinjection of the
pronucleus of the fertilized egg or transfection of ES cells with YACs
• carry either an array of related genes or a single large gene • used to study developmental processes • as models for human disorders • and for the production of human therapeutic agents.
YAC transgenesis • For example, transgenic mice with the human -globin gene cluster,
• covers about 250kb and contains five different functional globin genes,
• express genes in a tissue-specific and timedependent manner that corresponds to the site specificity and temporal pattern in humans.
YAC transgenesis • production of mice that synthesize only human antibodies
• Human monoclonal antibodies • Xenomouse
Schematic representation of the κ and H human immunoglobulin genes.
Transgenesis with High-Capacity Vectors • transgenes are complementary DNAs (cDNAs), small genes (<20 kb) or parts of genes
• cDNAs are poorly expressed in mammalian cells • In transgenesis, important gene- specific regulatory sequences retained as part of the insert
• • complete genes and multigene complexes (<100 kb) large for conventional vectors
• need for high-capacity vectors
• eg. bacterial, P1 bacteriophage- derived, mammalian, and yeast artificial chromosomes [ BACs, PACs,MACs and YACs, respectively])
• carry genomic DNA ranging in size from 100 to >1,000kb • used for transgenesis
YAC transgenesis • transgenic mice produced by microinjection of the
pronucleus of the fertilized egg or transfection of ES cells with YACs
• carry either an array of related genes or a single large gene • used to study developmental processes • as models for human disorders • and for the production of human therapeutic agents.
YAC transgenesis • For example, transgenic mice with the human -globin gene cluster,
• covers about 250kb and contains five different functional globin genes,
• express genes in a tissue-specific and timedependent manner that corresponds to the site specificity and temporal pattern in humans.
YAC transgenesis • production of mice that synthesize only human antibodies
• Human monoclonal antibodies • Xenomouse
Schematic representation of the κ and H human immunoglobulin genes.
