ANIMAL SCIENCESSCIENCE DEVELOPMENTS IN

TRANSGENIC ANIMAL ENGINEERING AND ASSISTED REPRODUCTIVE

TECHNOLOGIES

ADVISORY COMMITTEE ON BIOTECHNOLGY AND ADVISORY COMMITTEE ON BIOTECHNOLGY AND 2121STST CENTURY AGRICULTURE CENTURY AGRICULTURE

Biotechnologies, Animal Biotechnologies, Animal Production, and Animal HealthProduction, and Animal Health

1945 to 19951945 to 1995• Milk Production Up 3-Fold• Eggs/Year Up 134 to 254 • Broilers Days to 1.8 kg: Reduced from 84

to 43 Days and one-half the feed• Leaner and Faster Growing Pigs• Leaner and More Efficient Beef Cattle• Overall Improvement in Animal Health

Research Driven Biotechnologies in Research Driven Biotechnologies in Animal Agriculture and Biomedical Animal Agriculture and Biomedical

ResearchResearch

IMPROVED UNDERSTANDING OF:

Reproductive BiologyEndocrinology

EmbryologyGenetics/Genomics

NutritionAnimal Health/Disease Resistance

BIOTECHNOLOGIESBIOTECHNOLOGIESESTABLISHED: BREEDING AND GENETICS• Statistical Models Predict Breeding Values• Sire Testing and Selection• Crossbreeding

EMERGING• Marker Assisted Selection• Quantitative Trait Loci (QTL)/Single Nucleotide

Polymorphisms (SNPs)• Direct Gene Transfer • Homologous Recombination• Genetic Engineering Through Sperm

BIOTECHNOLOGIESBIOTECHNOLOGIESREPRODUCTIVE MANAGEMENT

ESTABLISHEDArtificial InseminationSynchronization of Estrus and OvulationSex Reversal (Fish)Artificial Incubation of EggsNutritionDisease Control

EMERGINGSEX CONTROLPREIMPLANTATION GENETIC DIAGNOSISTRAIT SELECTION

BIOTECHNOLOGIESBIOTECHNOLOGIESGAMETE AND EMBRYO MANIPULATIONS

ESTABLISHEDSuperovulation, Embryo Collection and Transfer,

IVM and IVF, Embryo Splitting and Cloning from Blastomeres

EMERGING Cloning

Somatic Cell Nuclear Transfer (SCNT)SCNT for Genetic Engineering

Epigenetics

Embryo SplittingEmbryo Splitting

IMPACT OF BIOTECHNOLOGIESIMPACT OF BIOTECHNOLOGIES

Assisted Reproductive Technologies/Cloning

Embryo Transfer: 40,000-50,000 Calves/Year

IVM/IVF: 4,000 Calves in 1996

Blastomere Nuclear Transfer Cloning (61 males and 126 females)

Embryo Splitting (754 males and 1,472 females)

Reference: NAAB 1996 and Holstein Associate 2001

BIOTECHNOLOGIESBIOTECHNOLOGIESIMPACTING LIVESTOCK INDUSTRY

Steroid Growth Stimulants

Bovine Somatotropin - Increase Milk Yield

EMERGINGSPERM-MEDIATED GENE TRANSFER ECTOPIC DNA CONSTRUCTS

MARKER ASSISTED SELECTION

GENOME SIGNAL PROCESSING –Systems Biology

EMERGING BIOTECHNOLOGIESEMERGING BIOTECHNOLOGIES

SPERM MEDIATED GENE TRANSFERVIRAL EXPRESSION VECTORS – SPERM UPTAKEDNA POSITIVE PIGS – 0 TO 88%DNA POSITIVE – mRNA POSITIVE – 41-100%EFFICIENT AND INEXPENSIVESTABLE INTEGRATION OF TRANSGENERANDOM INSERTION OF TRANSGENEADVERSE POSITION EFFECTS ON GENE

EXPRESSIONLETHAL EFFECTS OF EXPRESSION VECTOR

VL30 (50 ng) vs PCMVhGH (1000 ng)/1 million sperm

LAVITRANO ET AL. MOL REPROD DEV 64:284

EMERGING TECHNOLOGIESEMERGING TECHNOLOGIESEXPRESSION OF ECTOPIC DNA IN RATS

GHRH cDNA DRIVEN BY SYNTHETIC MUSCLE-SPECIFIC PROMOTER:

INCREASE GROWTH HORMONEINCREASE IGF-I

INCREASE FETAL-PLACENTAL DEVELOPMENTINCREASE BIRTHWEIGHTS

ALTERED PITUITARY SOMATOTROPHS AND LACTOTROPHS IN PUPS

KHAN ET AL. 2002; DRAGHIA-AKLI 2002

SPc5-12 hGH3’ kana

pSP-βgal

pSP-HV-GHRH

SPc5-12 HV-GHRH cDNA hGH3’ kana

β-gal

A.

Figure 1. RDA

B.

40

45

50

55

60

65

cf tf cm tm

wei

ght (

gram

s)

3 weeks

*

*

150

200

250

300

350

400

cf tf cm tm

wei

ght (

gram

s)

10 weeks

*

*

C.

0.2

0.3

0.4

0.5

0.6

0.7

0.8

cf tf cm tm

treatment group

mus

cle

wei

ght/

tota

l bod

y w

eigh

t *1

00

G/wt

TA/wt *

†

†

‡

3 weeks

0.02

0.12

0.22

0.32

0.42

0.52

0.62

cf tf cm tm

treatment group

mus

cle

wei

ght/

tota

l bod

y w

eigh

t*10

0

G/wt

S/wt§

§

24 weeks

SPc5-12 hGH3’ kanakana

pSP-βgal

pSP-HV-GHRH

SPc5-12 HV-GHRH cDNA hGH3’ kanakana

β-gal

A.

Figure 1. RDA

B.

40

45

50

55

60

65

cf tf cm tm

wei

ght (

gram

s)

3 weeks

*

*

40

45

50

55

60

65

cf tf cm tm

wei

ght (

gram

s)

3 weeks

*

*

150

200

250

300

350

400

cf tf cm tm

wei

ght (

gram

s)

10 weeks

*

*

150

200

250

300

350

400

cf tf cm tm

wei

ght (

gram

s)

10 weeks

*

*

C.

0.2

0.3

0.4

0.5

0.6

0.7

0.8

cf tf cm tm

treatment group

mus

cle

wei

ght/

tota

l bod

y w

eigh

t *1

00

G/wt

TA/wt *

†

†

‡

0.2

0.3

0.4

0.5

0.6

0.7

0.8

cf tf cm tm

treatment group

mus

cle

wei

ght/

tota

l bod

y w

eigh

t *1

00

G/wt

TA/wt *

†

†

‡

3 weeks

0.02

0.12

0.22

0.32

0.42

0.52

0.62

cf tf cm tm

treatment group

mus

cle

wei

ght/

tota

l bod

y w

eigh

t*10

0

G/wt

S/wt§

§0.02

0.12

0.22

0.32

0.42

0.52

0.62

cf tf cm tm

treatment group

mus

cle

wei

ght/

tota

l bod

y w

eigh

t*10

0

G/wt

S/wt§

§

24 weeks

EMERGING TECHNOLOGIESEMERGING TECHNOLOGIES

EXPRESSION OF ECTOPIC DNA IN PIGS

GHRH cDNA DRIVEN BY SYNTHETIC MUSCLE-SPECIFIC PROMOTER:

HEAVIER BIRTH WEIGHTSHEAVIER WEANING WEIGHTS

INCREASE SERUM IGF-IINCREASE IN PITUITARY SOMATOTROPHS AND

LACTOTROPHS (BIRTH AND 100 KG)

KHAN et al. 2003, Am J Physiol Endocrinol Metab 285:E224-E231.

IGF1 SNP 5A Allele and Size of Dog

Why Clone AnimalsWhy Clone Animals

Research

Development, aging, cancer, models for biomedical research

Applications

Genotype replication, genetic conservation, genetic engineering

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