8. Effect of the herbicide bromoxynil on tobacco plants transformed with a bacterial gene whose product breaks down bromoxynil (top row) and control plants (bottom row). "Spray blank" plants were treated with the same spray mixture as the others except the bromoxynil was left out. (Courtesy of Calgene, Davis, CA.)
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11. Direct microinjection • inject DNA molecules (transgenes) directly into male pronucleus • manipulated fertilized ovum is transferred into the oviduct of a recipient female, or foster mother • most popular technology, commercial available • the success rates range from 10-30% depending on skills and constructs • the insertion of DNA is a random process, and there is a high probability that the introduced gene will not insert itself into a site on the host DNA that will permit its expression • the efficiency is not related to the copies of transgenes injected • initial investment is high, a minimum of $100K to start
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13. Nuclear transfer • creation of Dolly • somatic cells be transfected, or genetically altered prior to NT • 100% efficiency of any progeny • abnormal development WHAT IS DOLLY????
17. KNOCK OUT MOUSE. What is a knockout mouse? A knockout mouse is a genetically engineered mouse in which one or more genes have been turned off through a gene knockout. Knockout mice are important animal models for studying the role of genes which have been sequenced, but have unknown functions. By causing a specific gene to be inactive in the mouse, and observing any differences from normal behaviour or condition, researchers can infer its probable function. Mice are currently the most closely related laboratory animal species to humans, for which the knockout technique can easily be applied.
18. The Nobel Prize in Physiology or Medicine 2007 "for their discoveries of principles for introducing specific gene modifications in mice by the use of embryonic stem cells“ is awarded to.. Mario R. Capecchi USA University of Utah Sir Martin J. Evans United Kingdome Cardiff University Oliver Smithies USA University Of North Carolina The first knockout mouse was created by Mario R. Capecchi , Martin Evans and Oliver Smithies in 1989 , for which they were awarded the Nobel Prize for Medicine in 2007 .
23. Targeted ES cells are inserted in to blastocyst of foster mother Chimeric F1 mice Chimeric male is crossed with normal female. Targeted (homozygous) & normal mice. (F2)
28. Monogenic Disease Lesch-Nyhan syndrome Cystic fibrosis inherited heart diseases Cancer Other diseases Obesity, Diabetes, Arthritis, Substance abuse, Anxiety, Aging and Parkinson's disease. Significance of gene targeting for physiology and medicine Knockout mice gives information that can be used to better understand how a similar gene may cause or contribute to disease in humans. Complex Disease Essential hypertension Atherosclerosis Atherosclerosis Inherited Heart Disease
29. Lesch-Nyhan syndrome , due to mutation in HPRT (phosphoribosyltransferase ) gene cause a defective nucleotide metabolism in human. Similar gene adenine phosphoribosyltransferase (APRT) for purine salvage in mouse was observed, cause of neuropathological & change in behavioral feature. Cystic fibrosis, Defective chloride transport through cAMP-activated chloride channel in mouse due to knocking out of cystic fibrosis transmembrane conductance regulator (CFTR) gene . The similar phenotypes is observed in human as in mice. Essential Hypertension, 10 genes are responsible for altering blood pressure. Angiotensinogen (AGT) gene polymorphism is associated with essential hypertension. Targeting of Angiotensin-converting enzyme (ACE) coding gene in mouse is observed to be effective in reduction of hypertension. Similarly renin-angiotensin system is applied in human for hypertension. Cancer, Protooncogenes, tumor suppressor genes, angiogenetic factors targeted in mice to know about the induction and spreading of tumours & their role in the formation of tumors. These targeted mouse being solid support to study of cancer in human. Gene Targeting In Disease Diagnosis And Study
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32. Interestingly, the creation of transgenic animals has resulted in a shift in the use of laboratory animals, from the use of higher-order species such as dogs to lower-order species such as microbes, and has decreased the number of animals used in such experimentation, especially in the development of disease models. This is certainly a good turn of events since transgenic technology holds great potential in many fields, including agriculture, medicine, and industry. CONCLUSION THANK YOU !