methods of forming transgenic mice and application of transgenic mice in field of biotechnology

1. PRESENTATION OF TRANSGENIC
MICE : METHODS AND APPLICATIONS
Made by- Teena
Roll no-5
Msc final year Biotechnology
Submitted to-Professor Anita yadav
Kurukshetra university

2. CONTENT
1.Introduction
2.Methods of generating Transgenic Mice
a)retroviral vector method
b)microinjection method
c)engineered embryonic stem cell
method
d)yeast artificial chromosome transgenesis
3.Applications of Transgenic mice
4.Refrences

3. Introductionof foreign DNA into the mouse
germ line is considered a major technical advancement in the
fields of developmental biology and genetics. This
technology now referred to as transgenic mouse technology
has revolutionized virtually all fields of biology and provided
new genetic approaches to model many human diseases in a
whole animal context. Several hundreds of transgenic lines
with expression of foreign genes specifically targeted to
desired organelles/cells/tissues have been characterized.
Further, the ability to temporally inactivate or activate gene
expression in vivo using the “Cre-lox” technology has
recently emerged as a powerful approach to understand
various developmental processes including those relevant to
molecular endocrinology.

4. Methods of generating
transgenic mice
1.Retroviral vector method
2.DNA Microinjection method
3.Engineered embryonic stem cell method
4.Clonning by nuclear transfer
5.Yeast artificial chromosome Transgenesis

6. Retroviral method

7. Microinjection method

9. Engineered embryonic stem cell
method

10. Positive–negative selection. (A) Result of nonspecific integration. Both genes for thymidine kinase (tk1 and tk2),
the two DNA sequences that are homologous to a specific chromosomal region in the recipient cells (HB1 and
HB2), a gene (Neor ) that confers resistance to the cytotoxic compound G-418, and the transgene (TG) are
incorporated into the chromosome. After transfection, cells are selected for resistance to both G-418 and the
compound ganciclovir, which becomes cytotoxic to cells that synthesize thymidine kinase. Other
nonhomologous integrations may occur and produce inserts with one or the other of the thymidine kinase
genes. After treatment with G-418 and ganciclovir, all the cells with nonspecific integration of the input DNA
that includes at least one of the thymidine kinase genes are killed. (B) Result of homologous recombination.
The product of a double crossover between homologous blocks (HB1 and HB2) of DNA on the vector DNA and
on chromosomal DNA does not contain either of the two thymidine kinase genes (tk1 and tk2). After treatment
with G-418 and ganciclovir, only cells that have undergone homologous recombination survive.

11. Cloning by Nuclear Transfer

12. Yeast Artificial Chromosome
Transgenesis

14. Transgenic Mice : Application
Transgenic mice can be used as
model systems for determining the
biological basis of human disease
and devising treatment for various
condition.

15. 1. Transgenic disease models: Alzheimer
Disease
Alzheimer disease have mutations in APP(beta- amyloid
precursor protein).A number of transgenic mice have
been generated with full or partial versions of the
normal APP gene driven by neurospecific promoters.
2.Tansgenic mice have also been used as
models for expression systems that are designed for
secretion of the product of a transgene into milk, For
eg: Large quantities of authentic cystic fibrosis
transmembrane regulator(CFTR) protein are needed to
study its function and formulate potential therapy for
treating cystic fibrosis.

16. 3.Using Transgenic mice as test system
The testing of RNAi in transgenic mice as a potential therapy
to reduce levels of protein that contibute to Alzheimer disease
in one of many example of transgenic laboratory animals in
testing strategies to treat human genetic diseases.
4.Conditional Control of Cell Death
The ability to induce cell death at different times and under
defined conditions in a specific organ, is a way to study organ
failure. Transgenic mice have been engineered for this
purpose. For e.g. to examine the effects of live cell damage,
transgenic mice were created to express a receptor that is
required for a bacterial toxin to cause cell death

17. REFRENCES
1.Molecular biotechnology: principles and applications of recombinant
DNA (Bernard Glick and Pasternak)4th Edition Chapter 21
2.Gurdon, JW and Ruddle, F.H(1981) Integration and stable germline
transmission genes injected into mouse pronuclei .Science, 214:1244-
1246
3.Houdebine, L.M(2002).The method to generate transgenic animals
and to control transgenic expression J.Biotechnol 98:145-160
4.Cornel, M.(2007): Molecular Biology and Genomics 1st edition,Elsevier
Inc,San diego Pg 116-118
5.Bagle , S.Kuknkulol,R; Baig ,M nad More, S.(2013):Transgenic animals
and their application in medicine Int. J.Medical Res. And Health Sci:2
107- 116
6. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4095860/ (this
enlist all the materials and steps required for transgenic mice
generation)