gene transfer

1. PRESENTED BY
“ BAKHTAWAR NAEEM,,
ASSIGNED BY “DR. YAAR
MUHAMMAD ,,

2. 👉USE OF BIOTECHNOLOGY
TECHNIQUES IN ANIMAL
BREEDING STRATEGIES
• GENE TRANSFER THROUGH EMBRYO MICROINJECTION

3. 👉WHAT IS GENE TRANSFER?
• Gene transfer is a process to transfer a gene from one DNA molecule to another DNA
molecule.
• It’s is simply defined as a technique to efficiently and stably introduce foreign genes into the
genome of target cells .
• Gene transfer technology provides the ability to genetically manipulate the cells of higher
animals. Gene transfer permits both germline and somatic alterations. Such genetic
manipulation is the basis for animal transgenesis goals and gene therapy attempts.

4. 👉CONTINUE
• This area of genetic manipulation makes important contributions to domesticated
animals in relation to immunology ,vaccine, aging and cancer .

5. 👉GENE TRANSFER THROUGH MICROINJECTION
• Introduction
• DNA microinjection was 1st proposed by Dr Marshall A. Barber in the early of 19th
century.
• This method is widely used for gene transfection in mammals .
• It involves delivery of foreign DNA into a living cell (e.g a cell , egg
oocyte,embryos of animals)through a fine glass micropipette.
• It is used to identify the characteristic function of dominant genes.

6. 👉APPLICATIONS
• Microinjection process is applicable for animal cells as well as
plants cells but more common for animals al cells.
• This technique is ideally useful for producing transgenic animals
quickly .
• Procedure is important for gene transfer to embryonic cells .

7. 👉ADVANTAGES
• Frequency if stable integration of DNA is far better as compare to other methods .
• No requirement of a marker gene .
• Intraditional of the target gene directly into a single cell .
• Easy identification of transformed cells upon injection of dye along with the DNA .
• Mere precise integration of recombinant gene in limited copy number can be
obtained.

8. 👉LIMITATIONS
• Costly
• The procedure is time consuming.
• Only a small number of cella can be treated .
• It requires specialised equipment and technical skills to prevent cell or DNA
damage.
• Result obtained by this method vary with the investigator’s experience in impaling
cells without causing too much damage .

9. 👉PROCEDURE
• Microinjection is performed by observing the recently fertilized
single-cell embryo through an inverted microscope.
• Differential interference contrast (DIC) optics are necessary to
enhance the contrast and provide clear visualization of the
embryo’s pronuclear structure.
• Microinjection of single-cell embryos is typically performed at a
magnification of 200×. Manipulation of the embryo is
accomplished using two micromanipulators and joystick
controllers located on either side of the microscope.
• One micromanipulator controls the movement of the holding
pipette, while the other controls the injection needle, both of
which are connected to finely controlled pressure systems to
allow fine control of the embryo and injection needle.

10. 👉
• Microinjection of foreign DNA into the pronucleus of a single-cell mouse or rat embryo is
accomplished by first picking up and securing the embryo with the holding pipette.
• It is generally accepted that microinjection into the male pronucleus (contributed from the
sperm) results in a higher rate of success in terms of transgenic offspring produced.
• The male pronucleus is generally the larger of the two pronuclei present and is typically located
at the periphery of the cell.
• The micromanipulator controlling the injection needle is then used to bring the needle into
sharp focus in the same focal plane as the pronucleus.
• The injection needle then carefully punctures the outer zona pellucid and cell membrane.
Injection is continued until the needle penetrates the pronuclear membrane and approximately
1–5 pL of the DNA solution is introduced to the pronucleus.

11. THANK YOU

12. ANY QUESTION ?