Scientists can now transfer genes across species, families and even kingdoms by HOW?
transferring to fertilized eggs or living cells of animals
Genetically engineered livestock will produce important products in blood and milk for health needs including... NAME 3
human hemoglobin, clotting agents, deficiencies
GENE TRANSFER METHOD OF CHOICE FOR MOST RESEARCHERS.
WHAT AND WHEN?
Pronuclear microinjection (1980)
Pronuclear microinjection WHAT TYPE OF MICROSCOPE?
Differential interference contrast microscope
3 PROBLEMS WITH GENE TRANSFER METHOD
1. low survival of cells
2. low efficiency of homologous recombination
3. gene targeting in animals
4. gene usually inserts itself into the chromosomes at random
THE STEPS OF GENE TRANSFER.
7 OF THEM
Identification gene of interest
Insertion foreign gene into an appropriate vector
Microinjection of DNA into the pronucleus of EGG
Implantation egg into a surrogate mother
Develop to birth
Demonstrating stably incorporated to offspring
Demonstrating that the gene is expressed and regulated correctly in the host organism
DEFINE Pluripotent
Capable of differentiating into other cell types
Gene Transfer Methods mediated gene transfer into germ line cells (reproductive)
NAME 5
RETROVIRUS
EMBRYONIC STEM CELLS
Electroporation,
virus infection,
DNA precipitation onto cell surface
(calcium phosphate for competence)
DEFINE TRANSFECTION
INSERTING A GENE TO BE TRANSFERED INTO ES CELLS. BEFORE THEY GET MICROIINJECTED INTO THE BLASTOCYST.
DEFINE KNOCKOUT
Insertion of DNA into a specific chromosomal location; Often used to inactivate OR KNOCKOUT a gene.
The knockout mice are confirmed for
the target gene with a DNA test like ------
PCR
DEFINE HOW TO KNOCKOUT A GENE.
1. START WITH ES CELLS THAT R HETEROZYGOUS FOR "KNOCKOUT" BLACK COAT. (NONFUNCTIONAL)
2. INJECT EMBRYO WITH BOTH NON AND FUNCTIONAL
3. BREAK OFFSPRING WITH HETERO
4. NEXT GEN ALL BLACK
---------- ------- GENE is used and will activate a toxic
substance if randomly inserted into the host chromosome (use ganciclovir as drug)
Thymidine kinase
T OR F?
Only cells that undergo homologous recombination survive treatment with antibiotic
and ganciclovir (PCR can be used to screen)
TRUE
Gene targeting can be a very means of creating animal models for human diseases
TRUE
-------- human diseases are caused by genetic defects
5,000
Cytic fibrosis transmembrane regulator (CFTR) is being studied and manufactured WHERE?
murein mammary glands (milk fat globules)
Cows are given specific traits by
microinjections of genes into eggs.
NAME 3
Over expression of k-casein gene, or lactose free
Increased resistance to pathogens like mastitis
Recombinant bovine somatotropin (more milk)
Pigs,Sheep and goats
NAME MICROINJECTION PRODUCTS
Pig...BIOREACTORS
Sheep and goats:
production of human proteins in milk
clotting factor VIII and IX, growth hormone, and interleukins
ENGINEERED BIRD PRODUCTS
Made resistant to virus and bacteria
Poultry and eggs made with less fat or cholesterol
Eggs could be used as factories for producing valuable human proteins
NAME 2 ISSUE WITH BIRDS AND MICROINJECTION.
-instability of gene and size restriction of DNA inserts
-does not work because of the encasement SHELL
-------- -------- & ---------- --------- that are low cost and eliminates the use of attenuated viral or bacterial pathogens
Monoclonal antibodies
recombinant vaccines
Foot and mouth disease Extremely contagious,
------ virus particles to infect a cow
10
INACTIVED VACCINE, NEED REFRIDG.
NO LONG TERM PROTECTION.
STRAIN SPECIFIC
Believed to be caused by a prion, which is a poorly understood agent that may be an
infectious protein
MAD COW.
Bovine spongiform encephalopathy (BSE)
------- are very resistant to sterilization and do not evoke a detectable immune response or inflammatory reaction in hosts, and no treatments are available
PRIONS
Humans can be infected with a new variant called ----------- which causes nervous system tissue to look sponge-like
AND WHAT DIESASE ?
Creutzfeldt-Jakob disease (nvCJD)
MAD COW
Bovine spongiform encephalopathy (BSE)
DEFINE Coccidiosis
parasitic protozoans in the genus Eimeria that invade the epithelial cells of the digestive tract and associated glands in cattle, sheep, and poultry.
CROWDED CONDITIONS.
MANY FEEDS CONTAIN ANTICOCCIDIAL DRUG
VACCINE..TRIAL..DESTROY OOCYSTS
DEFINE Trypanosomiasis
-One of the most studied parasites; African sleeping sickness.
-parasite changes its surface antigen during infection
and escapes detection
Trypanosomiasis. Research has focused on three areas:
- trypanosome components involved in the process (VACCINES)
-Isolating resistance genes IN AFRICAN cattle
-Host genes that inhibit parasite division
DEFINE Theileriosis
East Coast fever..... cattle disease .....ticks
infects lymphocytes, which become leukemic and
cause lymphosis, killing the cattle.
Use of monoclonal antibodies or sporozoite antigens for vaccination
WHAT DISEASE?
Theileriosis
Diluted sperm sample from one bull
will inseminate _____ - _______ females
500-1000
Produce identical twins (cattle breeding) two superior animals from one ,Increases the rates of specific
traits.
KNOWN AS WHAT?
EMBRYO TWINNING
First successful animal cloning was of a sheep in ----
Mice were first cloned in -----
FROG STUDY--------
1996
1998
1950'S
Nuclear transfer methods:
Uses ------- ----- FROM ----- -----.
pluripotent nuclei from embryonic cells
Totipotency
Ability of a single cell to divide and produce all the differentiated cells in an organism, including extraembryonic tissues, include spores and zygotes. In some organisms, cells can dedifferentiate and regain totipotency.
1996, Dolly the sheep cloned using
nucleus from a differentiated cell from ------ tissue.
mammary
Dolly died in ---- of ------- ------.
20003 of lung cancer
DOLY'S DNA analysis showed that the ends
of the chromosomes called ----------- were shorter than normal
TELOMERS
Controversy remains; it is not clear if Dolly was cloned from a mammary cell or from a stem cell present in
--------- ------- ------- ------.
the udder of the donor sheep
IT took ----- attempts to clone
Dolly, which is very inefficient
277
Genetic diversity preservation.
NAME 4
-Advances in in vitro fertilization, embryo transfer,
oocyte maturation, control of ovarian cycle
-captive breeding programs
-Cryopreservation of gametes and
embryos..... frozen germplasm
-DNA fingerprinting can be used for ID of animals and to resolve genetic differences
EMBRYO TRANSFER IS DONE AT WHAT TEMP?
Fertilized eggs are retrieved from female and frozen at -196ºC and implanted into surrogate females made
pseudo-pregnant
Cincinnati Zoo has had success using Holstein cows to carry rare species..WHAT SPECIES?
Malaysian ox or Gaur, or the bongo antelope using Elands
Bongos from Mt. Kenya were implanted with bongo embryos from the US...WHY?
meant to increase genetic diversity in the wild and is closely monitored
In 2003, legislation called the Minor Use
and Minor Species Animal Health Act was introduced in Congress to DO WHAT?
improve the access of farmers to animal drugs.
ISSUE: FDA WILL REGULATE ANIMALS LIKE DRUGS.
Plant patents have been around
since the ------- (asexually propagated).
-------- sexually propagated plants
were made patentable.
--------US Supreme Court made landmark decision concerning patenting mammals.
1930'S
1970
1980
Non-naturally occurring non-human multicellular organisms were made patentable....WHEN AND WHO?
1987 by Don Quigg
patent # 4,736,866 was granted for a genetically altered mouse, was used to study human breast cancer was the first mammal patented
WHO AND WHEN?
1988 HARVARD
Cloned animals problems.
NAME 5
-die soon after birth
-low success efficiency,
-large offspring syndrome
-physical deformities
-Arthritis, heart disease, and respiratory problems are also common.
