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Genetic EngineeringSearching for a target for the treatment ofatherosclerosis
RISK FACTORSConsumption of foods high in saturated fats and a sedentary lifestyle increase therisk of suffering from cardi...
WHAT IS ATHEROSCLEROSIS?Atherosclerosis is a vascular disease caused by the accumulation of fats on thewalls of the blood ...
“BAD” OR LDL CHOLESTEROL                           • When there is too much cholesterol                           in the d...
ACCORDING TO THE WORLD HEALTH ORGANISATION (WHO)•Although heart attacks and strokes are major killers in all parts of the ...
THE MACROPHAGES, A “CLEAN-UP SYSTEM”• The macrophages act as a “cleaning” system to stop the cholesterol from beingdeposit...
HOW DOES AN ATHEROMATOUS PLAQUE FORM?                                                                                 Prol...
RESEARCH INTO ATHEROSCLEROSISOne of the objectives is:To understand how macrophages are involved in the regulation of chol...
STUDY OF THE LDL RECEPTOR AND MYLIPWe know that the macrophages recognise oxidised LDL thanks to some receptors.We know th...
MYLIP, A POTENTIAL THERAPEUTIC TARGETScientists are studying the MYLIP protein in depth because they think that it could b...
HOW CAN WE STUDY THE PROTEINS?• Large quantities of a protein are needed tostudy its function.•One of the tools in molecul...
FIRST WE CLONE THE GENE OF THE PROTEIN OF INTEREST:MYLIP                                     We insert it into a fragment ...
WE PERFORM A BACTERIAL TRANSFORMATION WITH THEGENE OF THE PROTEIN OF INTEREST: MYLIP1. Bacterial     2. Selection of trans...
THE GENE OF THE MYLIP PROTEIN IS THEN INTRODUCEDINTO CELLS IN ORDER TO PRODUCE MORE                                       ...
HOW DO WE DO A BACTERIAL TRANSFORMATION?                        With drastic changes in       ?                temperature...
HOW CAN WE MAKE SURE THAT TRANSFORMED BACTERIAWILL GROW?                    ?The plasmid has a genethat is resistant to an...
HOW CAN WE MAKE SURE THAT TRANSFORMED BACTERIAWILL GROW?
HOW DO WE ISOLATE THE PLASMID DNA FROM THEBACTERIAL CULTURE?                                                         ?Doin...
WITH THE DNA ISOLATED, WE CAN GET ON WITH THERESEARCH!                     5. Production of the protein: (introducing the ...
Put on a lab coat!!                                                                              You too can be a research...
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Genetic engineering - Searching for a target for the treatment of atherosclerosis

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Protocol for youngsters to carry out a bacterial transformation in a lab. The protocol follows a line of biomedical research which focuses on the study of a potential therapeutic target that could be recognised by a drug against atherosclerosis. The experiment protocol is an opportunity for science centres, museums and schools to replicate a real experiment done in a real lab doing research on drug discovery.

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Transcript of "Genetic engineering - Searching for a target for the treatment of atherosclerosis"

  1. 1. Genetic EngineeringSearching for a target for the treatment ofatherosclerosis
  2. 2. RISK FACTORSConsumption of foods high in saturated fats and a sedentary lifestyle increase therisk of suffering from cardiovascular diseases such as atherosclerosis.
  3. 3. WHAT IS ATHEROSCLEROSIS?Atherosclerosis is a vascular disease caused by the accumulation of fats on thewalls of the blood vessels. There are many different signs and degrees of severity.Normal arteryModerateatherosclerosisSevere atherosclerosis Formation of atheromatous plaque, which obstructs the blood flow.
  4. 4. “BAD” OR LDL CHOLESTEROL • When there is too much cholesterol in the diet, it either ends up being stored as fat or continues circulating in the blood in the form of LDL (low- density lipoprotein) or what is more commonly known as “bad cholesterol”. • LDL is one of the components of the atheromatous plaque.
  5. 5. ACCORDING TO THE WORLD HEALTH ORGANISATION (WHO)•Although heart attacks and strokes are major killers in all parts of the world,80% of premature deaths from these causes could be avoided by controlling themain risk factors•Every year, an estimated 17 million people globally die of cardiovasculardiseases (CVD), particularly heart attacks and strokes.•CVDs occur almost equally in men and women.•Are the leading cause of death in developing countries, as well as developedones.
  6. 6. THE MACROPHAGES, A “CLEAN-UP SYSTEM”• The macrophages act as a “cleaning” system to stop the cholesterol from beingdeposited on the walls.• This clean-up system is efficient if the increased cholesterol is not too excessive. LDL Macrophage Oxidised LDL Oxidation of LDL
  7. 7. HOW DOES AN ATHEROMATOUS PLAQUE FORM? Proliferation of endothelialIf the quantities of cholesterol are very cellsexcessive: Immune system activation The macrophages continue to pick up the LDL Foam But, once they have engulfed large amounts, they turn cell into what is known as “foam” cells. LDL MacrophageThese induce inflammation and the proliferation of cells in the artery wall Oxidised LDL Oxidation of LDL Formation of the atheromatous plaque
  8. 8. RESEARCH INTO ATHEROSCLEROSISOne of the objectives is:To understand how macrophages are involved in the regulation of cholesterol levels and what role they play inthe development of atherosclerosis. ?
  9. 9. STUDY OF THE LDL RECEPTOR AND MYLIPWe know that the macrophages recognise oxidised LDL thanks to some receptors.We know that a protein in the macrophages called MYLIP breaks down the LDLreceptor. Macrophage LDL receptor Oxidised LDL If the macrophages produce MYLIP in large quantities, they ingest less Oxidation of LDL cholesterol.
  10. 10. MYLIP, A POTENTIAL THERAPEUTIC TARGETScientists are studying the MYLIP protein in depth because they think that it could beregulated by a drug and thus stop the macrophages from ingesting less cholesterol. Macrophage Therapeutic target Oxidised LDL Oxidation of LDL
  11. 11. HOW CAN WE STUDY THE PROTEINS?• Large quantities of a protein are needed tostudy its function.•One of the tools in molecular biology whichallows us to study the proteins is geneticengineering.•Genetic engineering is the technology of themanipulation and transfer of DNA from oneorganism to another.
  12. 12. FIRST WE CLONE THE GENE OF THE PROTEIN OF INTEREST:MYLIP We insert it into a fragment of Human cell ADN MYLIP gene circular DNA called plasmid
  13. 13. WE PERFORM A BACTERIAL TRANSFORMATION WITH THEGENE OF THE PROTEIN OF INTEREST: MYLIP1. Bacterial 2. Selection of transformed 3. Bacterial growthtransformation bacteria
  14. 14. THE GENE OF THE MYLIP PROTEIN IS THEN INTRODUCEDINTO CELLS IN ORDER TO PRODUCE MORE Protein MYLIP 3. Bacterial 4. We isolate 5. Production of growth the DNA the protein (introducing the DNA into eukaryotic cells)
  15. 15. HOW DO WE DO A BACTERIAL TRANSFORMATION? With drastic changes in ? temperature and by adding cations, we help the entry of the DNA in the form of plasmid into the cells.
  16. 16. HOW CAN WE MAKE SURE THAT TRANSFORMED BACTERIAWILL GROW? ?The plasmid has a genethat is resistant to anantibiotic.
  17. 17. HOW CAN WE MAKE SURE THAT TRANSFORMED BACTERIAWILL GROW?
  18. 18. HOW DO WE ISOLATE THE PLASMID DNA FROM THEBACTERIAL CULTURE? ?Doing a Mini-prep:Using various solvents and centrifugation cycles, we gradually discard thedifferent components
  19. 19. WITH THE DNA ISOLATED, WE CAN GET ON WITH THERESEARCH! 5. Production of the protein: (introducing the DNA into eukaryotic cells). 6. Once the scientists obtain the protein, they can then: • Study its role in cholesterol regulation • Search for new drugs for atherosclerosis.
  20. 20. Put on a lab coat!! You too can be a researcher!! Researchers who have contributed to the writing of this presentation: Theresa León, Jonathan Matalonga, Barcelona UniversityAUTHOR FUNDED BY: PROJECT PARTNERS: This work is under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported licence. To see a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/3.0/
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