1. Transgenesis with high capacity vectors
Lecture 26
Transgenesis with high capacity vectors

2. 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

3. 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

4. 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.

5. 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.

6. YAC transgenesis
production of mice that synthesize only human antibodies
Human monoclonal antibodies
Xenomouse

7. Schematic representation of the κ and H human immunoglobulin genes.