Transgenesis with high capacity vectors 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 time- dependent 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.