Gene transfer methods in animalsPresentation Transcript
Activity: Identify set of pictures andcombined all of their first letters. Tie
Gene Transfer The insertion of unrelated genetic information in the form of DNA into cells. •Primary DNA •Secondary DNA
PlasmidSmall circular DNA molecule that replicates independently of the chromosome. •Origin of Replication •Antibiotic Resistance •Multiple Cloning Site
Transgenic organismAn individual with a genetic modification in every cell.
Microinjection process of introducing minute amounts of a substance into a microscopic structure, such as a single cell.
Direction:Arrange the following steps (A-F)• ___ Tissue samples from offspring are subjected to DNA analysis• ____ Pronuclei of male and female are separated• ____ Fertilized eggs from super ovulated will be removed and placed in sterile dish with buffer• ____ Implantation of microinjected egg to surrogate mother• ____ Identification of foreign gene of interest
Identification of foreign gene of interestFertilized eggs from super ovulated will be removed and placed in sterile dish with bufferPronuclei of male and female are separated DNA is injected to pronuclei Implantation of microinjected egg to surrogate motherTissue samples from progeny are subjected to DNA analysis
Animals with new gene injected into their germ cell line and somatic cells.
Embryonic stem (ES) cells Gene Transfer
Embryonic stem (ES) cells Cultured cell lines derived from the inner cell mass of the blastocyst
Early Mammalian Development
Embryonic stem (ES) cells
Embryonic Stem Cell Gene Transfer Totipotent Pluripotent
Direction:Arrange the following steps (A-D)• ___ Transformed ES cells are microinjected into animal blastocysts• ____ Es cells are culture in vitro• ____ Isolation of ES cells from inner cell mass• ____ Transfection of specific gene to ES cells
Isolation of ES cells from inner cell mass Es cells are culture in vitro Transfection of specific gene to ES cellsTransformed ES cells are microinjected into animal blastocysts
Gene TargetingIt uses a natural process calledHomologous Recombination inwhich a DNA sequence replaces asimilar or identical sequence in ahost chromosome.
Gene TargetingMicroinjection of inactivated gene into ES cell.Altered ES cells were injected into early embryos.Embryos were implanted into surrogate mothers
Gene Targeting in Mice
Retrovirus and gene transferRemoval of the retroviral genes forreverse transcriptase, protease, andintegrase are replaced by a therapeuticgene.