i have included terminology, types, methods, process, applications of trangenic technology. all the pics are collected from different websites and some text books shown in reference. pictures and matter copyrights doesn't belong to me.

1. TRANSGENIC ANIMALS
By
Konjeti. Surabhi.
620209504010,
A. U. pharmaceutical sciences.

2. Hai!
Welcome to
transgenic world

3. CONTENTS
Δ Introduction.
Δ History.
Δ Types.
Δ Process of transgenesis.
Δ Uses.
Δ Products and their production process.
Δ Recent advancements.
Δ Conclusion.
Δ Reference.

4. INTRODUCTION
• A transgenic animal is one whose genome has been altered by the transfer of a
gene or genes from another species or breed.
• Official term: GMO’s
• For Unicellular organisms (such as bacteria or yeast):
all transformed cells are -> transgenic
• For multicellular organisms (such as animals, plants,..) difference : -
manipulation of single cells -> cell line (expression in insect cells or
mammalian cells)
manipulation of a whole plant or animal -> transgenic (can have a transgenic
offspring!!!)

5. Terminology
◊ Transgene: the foreign DNA that is introduced.
◊ Transgenesis : a process where mixing up of genes occurs.
◊ Transgenic : the organism that develops after a successful gene transfer.
◊ Chimeric animal: Chimeras are animals composed of cells that originate from
two (or more) different species.
◊ Homozygous :having two identical alleles of a particular gene or genes.
◊ Transfection is the process of deliberately introducing naked or purified nucleic
acids into eukaryotic cells.

7. HISTORY
• 1891: Successful transfer of embryo byWalter Heape in Angora rabbits.
• 1940: In vitro manipulation of embryos in mice using culture system.
• 1966:Teh Ping Lin outlined a technique to micro-inject fertilised mouse eggs.
• 1974: Rudolf Jaenisch created a transgenic mouse by introducing foreign DNA
into its embryo, making it the world's first transgenic animal.
• 1976: retrovirus-mediated transgenesis (Jaenisch).
• 1981: first transgenic mouse(Gordon and Ruddle).
• 1982: first supermouse having GH (Gossler et al).
• 1987: First animal to synthesize transgenic proteins in their milk.

8. TYPES
Category Name specality example
First Disease models investigating disease
mechanisms and
potential cures
Alzheimer’s mouse,
AIDS Mouse, and
OncoMouse.
Second Transpharmers new sources for drugs
synthetically produce
Baby Herman,
Genzyme goats.
Third Xenotransplanters produce organs
compatible with
humans.
Pig Xenotransplanters
and baboon
Xenotransplanters
Fourth Transgenic food sources to produce more meat Superpig and Superfish
Fifth Scientific models In-vivo studies of novel
proteins(drugs)
Doogie

9. Process ofTrangenesis
• DNA
synthesis
Step 1
• Insertion of
transgene
Step 2 • Screening
of
genotype
Step 3
• Establishment
of F2
generation
Step 4

10. DNA Synthesis

11. Methods
• Pronuclear micro- injection
• Embryonic stem (ES) cell manipulation
• Viral vectors
• Nuclear transfer
• Cytoplasmic injection
• Cre-lox technique
• Spermatogonial manipulation
• CRISPR/Cas9

13. Pronuclear micro- injection
• Microinjection is a technique in which recombinant DNA is directly injected into the
nucleus of an animal. In this, through a glass micropipette(0.1 to 10µm), foreign
DNA is delivered directly into a living cell, oocyte or embryos of animal.
• Gordon and Ruddle, 1981.
• It is used to identify the characteristic function of dominant genes.
• Mostly suitable for mammals only.
• Highly reproducible and repeatable method of introducing genetic material into a
nucleus.
• Transgene integration into the genome of founder animals is low.

15. Embryonic stem cells
• Embryonic stem cells (ES cells) are harvested from the inner cell mass (ICM) of
mouse blastocysts.
• They can be grown in culture and retain their full potential to produce all the cells
of the mature animal, including its gametes.
• Out of the total progeny derived from such embryos, nearly 30% contain tissues
derived from the embryonic stem cells, and they are chimeric.
• Important for the study of the genetic control of developmental processes.
• It has the advantage of allowing precise targeting of defined mutations in the
gene via homologous recombination.

18. Viral vectors
• Retroviruses are commonly used as vectors to transfer genetic material into the
cell.
• Transmission of the transgene is possible only if the retrovirus integrates into
some of the germ cells.
• F1 generation may result in chimeras.
• Inbred until homozygous transgenic animal is produced.
• The embroys of these are frozen and stored for subsequent implantation.
• Efficient mechanism of transgene integration.

20. Nuclear transfer Cytoplasmic injection

21. Cre-lox technique ATAACTTCGTATA -NNNTANNN-TATACGAAGTTAT

22. Spermatogonial manipulation

24. CRISPR/Cas9

25. Somatic cell nuclear transfer

26. SCREENING OF GENOTYPE:
 Collection of DNA sample.
 Methods used in screening:
 RT-PCR - detects the RNA coding for trans-protein.
 ELISA - amount of trans-protein.
 Southern blot test – to det the no. of transgenic copies integrated.
 Western blot test - detects the proteins made from transgene.
 Some transgenes may not be expressed if they are inserted in other
than transgenic sites.

27. PROCESS OFTRANGENESIS
• DNA
synthesis
Step 1
• Insertion
of
transgene
Step 2 • Screening
of
genotype
Step 3
• Establishment
of F2
generation
Step 4
• Production of
homozygous
breed.
Step 5

28. PROBLEMS
• Insertion shouldn’t always be as the desired.
• Multiple insertions may occur.
• Not all insertions are fruitful.
• Gene regulation me be effected.
• Produce unnecessary problems to host.

29. LIST OF PURPOSES
• In Medical research, to identify the functions of specific factors in complex
homeostatic systems through over- or under-expression of a modified gene
(the inserted transgene).
• InToxicology: as responsive test animals (detection of toxicants).
• In mammalian developmental genetics.
• In Pharmaceutical industry, targeted production of pharmaceutical proteins,
drug production and product efficacy testing.
• In Biotechnology: as producers of specific proteins, enzymes etc.

30. Schematic representation of products produced using transgenic technology

31. Production Of Protiens

32. PRODUCTION OF PHARM ANIMALS

33. Production of rhAT milk

34. Production of mAB

35. Recent advancements
• Liu et al. produced transgenic chickens that human neutrophil defensin 4
proteins were successfully expressed in the chickens' eggs.
• Park et al. could genetically transform chickens for producing human
epidermal growth factor which was expressed by an oviduct-specific
promoter.
• Delivery efficiency of CRISPR/Cas9 elements to target locus and obtaining
sustained expression of the CFTR transgene to treat autosomal cystic
fibrosis of lungs.
• Fragment fused factor IX (FIX-Fc) in the milk of transgenic dairy animals.

36. CONCLUSION
• Transgenic technology is a field that is under constant evolution.
• All of the developments in transgenic technology will provide new ideas and bring
forth important changes in fields like medicine, health and livestock improvement.
• In particular, the economic and social benefits from the production of bioreactors,
drug production, and organ culture for human transplantation will be great.
• With proper research and careful use the transgenic animals can go a long way in
solving several problems for which science doesn't have a solution till now.
• The issue is where you draw the line between human benefit and animal
discomfort because ethics plays a major role in our field to gain what we want.

37. REFERENCE
• Molecular Biotechnology: Principles and Applications of Recombinant DNA, 4th
Edition Bernard R. Glick, Cheryl L. Patten pg.no: 845 -896.
• Pharmaceutical Biotechnology Drug Discovery and ClinicalApplications Second,
Completely Revised, and Greatly Enlarged Edition by Oliver Kayser and Heribert
Warzecha pg.no: 71-105.
• Renald Blundell Department of Physiology and Biochemistry :Journal of Animal and
VeterinaryAdvances 5 (11): 935-938,2006 © Medwell Journals, 2006 Transgenic Animals-
Review Paper.
• MiguelA. Gama Sosa • Rita De Gasperi • GregoryA. Elder :Brain Struct Funct (2010)
214:91–109 DOI 10.1007/s00429-009-0230-8 Animal transgenesis: an overview.
• Singhal & Kansara (2010),Vol.1 (9): 12-22 ISSN: 0975-8232 Available online on
www.ijpsr.com 12 IJPSR (2010),Vol. 1, Issue 9 TRANSGENICANIMALS: PRODUCTION
AND APPLICATION.

38. • https://www.scribd.com/document/452484850/Transgenic-Animals-and-Their-
Applications
• https://web.wpi.edu/Pubs/E-project/Available/E-project-081804-
222235/unrestricted/IQP.pdf
• https://biochemie.medizin.uni-
leipzig.de/cloning_club/images/stories/cloning_club_transgenic_animals.pdf
• https://studylib.net/doc/5785537/transgenic-animals---lungeninformationsdienst
• http://bcas.du.ac.in/wp-content/uploads/2020/04/Transgeni-Technology.pdf
• https://www.pashudhanpraharee.com/production-application-of-transgenic-
animal-for-improving-livestock-productivity-human-welfare/