Animal Biotechnology Lectures

Biotechnology, Biology 395

Spring 1998

10,12 March 1998

Kenneth L. Campbell

I) Transgenic Animals

A) Examples of uses
1) Nutrient sources
a) Fish -- salmon/growth hormone

b) Tomatoes -- Flavor Saver -- ripening arrest/shipping

c) Soy beans -- disease resistance

d) Strawberries -- frost resistance

2) Pharmaceutical production
a) human hormones from goat or cows milk, urine

b) antibiotics

3) Disease models
a) knockouts and reverse genetics

b) inherited recessive conditions

c) acquired or degenerative conditions nonendemic to model

B) Temporary transfection and expression
1) viral transfections

2) embryonic injection

3) tissue injection

4) somatic expression

C) Permanent transformation and expression
1) germ line transfection

2) pronuclear injection, developmental dynamics, chimeric transgenics

3) embryonic stem-cell transformation, selection, chimeras

4) episomes, random insertions, homologous recombinations

a) regulation of expression

b) timing

c) tissue specificity

II) Cloned mammals

A) Examples and issues
1) Dolly -- Wilmut, Campbell et al. -- adults from adults?
a) developmental expression programs

b) totipotency

c) telomerase

d) mitochondrial contributions

2) Calves -- UMA -- bioreactors for clotting factors
3) Rhesus -- Oregon -- Wolf et al. -- animal models/organ donors
4) Humans -- Is it possible?
B) Methods
1) Nuclear replacement in meiosis II ova by
a) embryonic cell nuclei

b) adult? cell nuclei from starved cells G0 arrest

c) reactivation via electronic pulse

2) Screening
a) unlinked genes

b) linked genes, markers

c) Southern blots

d) PCR

3) Breeding to homozygosity & generation of lines

Is metazoan transgenesis possible in birds or amphibians? What alterations in current mammalian methods would be needed to accommodate differences in physiology? Work out a routes to amphibians that are transgenic for loci on an autosome or on a sex chromosome.