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Recombination DNA Technology (Microinjection)
- 1. Subject :rDNA Technology SubjectCode:YBT201 Topic: Microinjection Submitted To, Ms.P Mala Assistant Professor Department of Biotechnology Periyar maniammai university Submitted by, Name: J.shifa Reg no:123011356017 Class:1st Msc, Biotechnology
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- 3. Contents Introduction toGene Transfer Methods of gene transfer Microinjection Principle of Microinjection Procedure Applications Disadvantages
- 4. Introduction to GeneTransfer It is defined simply as a technique to efficiency and stably introduce foreign genes into the genome of targeted cells. The insertion of unrelated ,therapeutic genetic Information in the form of DNA into targeted cells.
- 5. Methods of genetransfer There are two methods of gene transfer Natural method of gene transfer Artificial method of gene transfer
- 6. Methods of DNATransfer
- 7. Microinjection Microinjection isone of the physical methods of gene transfer used to introduce DNA or other genetic materials directly into a cell using a small glass needle or micropipette. Dr. Marshall A. Barber introduced the concept of DNA microinjection during the early 19th century. Since then, it has continually evolved to keep up with the advancements in biomedical fields This method allows efficient transfer and integration of desired genes into the host cell’s genome
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- 9. Principle of Microinjection The principle of microinjection is based on the direct delivery of genetic material into individual cells using a fine glass needle called a micropipette, a positioning device known as a micromanipulator, and a microinjector. Nikon’s ECLIPSE Ti2 and ECLIPSE Ts2R series of inverted microscopes, and SMZ18/25 stereo microscopes, are stable and vibration-resistant systems that can be used for microinjection work. The genetic material is delivered into the cell by applying hydrostatic pressure to release a fluid containing the DNA through the micropipette.
- 10. Microinjection procedure Thedelivery of foreign DNA is done under a powerful microscope Using a glass micropipette tip of 0.5mm diameter. Cells to be microinjected are placed in a container . A holding pipette is placed in the field of view of a microscope that sucks and holds a targeted cell at the tip. The tip of micropipette is injected through the membrane of the cell to deliver the Content of the needle into the cytoplasm and then the empty needle is taken out.
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- 12. Use of Transgenicmice in aging research Method of producing transgenic Mice by Microinjection
- 13. Applications of microinjection Microinjection is commonly used in the production of transgenic animals, introducing foreign DNA into fertilized eggs to study gene function and create disease models. Microinjection is also used in somatic cell nuclear transfer (SCNT) to create genetically identical copies of an organism by transferring a somatic cell into an enucleated oocyte. Microinjection has been widely used in various studies, including human embryo research and injection of mitotic cells.
- 14. Disadvantages Time consumingprocedure. Low success rate. Only one cell is targeted per injection Costly method and requires high skills.
- 15. MCQ Questions 1.Who introducedthe concept of DNA Microinjection? a.Dr. Marshall A. Barber b.Robert Hook c.Herbert Boyer 2.Microinjection is commonly used in the production of ________. a.Transgenic plants b.Transgenic Animals c.Both 3.The genetic material is delivered into the cell by applying _______ 2.________ is one of the physical method of gene transfer. 5.Microinjection is also used in ________ to create genetically identical copies of an organism .
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- 17. References Arlan Richardson,Ahmad R. Heydari, William W. Morgan, James F. Nelson, Z. David Sharp, Christi A. Walter, Use of Transgenic Mice in Aging Research, ILAR Journal, Volume 38, Issue 3, 1997, Pages 124– 136, Blanco, J. Transgenic animals. Worcester Polytechnic Institue. 2012. Ghaemi, R.; Tong, J.; Gupta, B.P.; Selvaganapathy, P.R. Microfluidic Device for Microinjection of Caenorhabditis elegans. Micromachines 2020, 11, 295. https://doi.org/10.3390/mi11030295 Lee, S.; Jeong, W.; Beebe, D.J. Microfluidic valve with cored glass microneedle for microinjection. Lab A Chip 2003, 3, 164–167.
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