Genetic engineering

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Dental aspects of genetic engineering is also covered.

Text of Genetic engineering

  • 1. GENETICENGINEERINGBy,Dr. Priyanka SharmaII year MDSDepartment of Public Health DentistryJSS Dental College & Hospital 11

2. CONTENTS1) INTRODUCTION2) BASICS OF GENETIC ENGINEERING3) HISTORY OF GENETIC ENGINEERING4) GENERAL APPLICATION OF GENETIC ENGINEERING5) GENETIC ENGINEERING IN DENTISTRY- VARIOUS TECHNIQUES OF GENETIC ENGINEERING6) GENETIC COUNSELLING & ROLE OF DENTIST7) SUMMARY8) CONCLUSION9) REFERENCES22 3. CONTENTS33 4. INTRODUCTIONGenetic engineering is a part ofbiotechnology.Biotechnology is the use of livingsystems and organisms to develop ormake useful products, or "anytechnological application that usesbiological systems, living organisms orderivatives thereof, to make or modifyproducts or processes for specific use"(UN Convention on BiologicalDiversity, Art. 2).44 5. INTRODUCTION continuation.. Biotechnology is a huge topic. Its hard to define its exact boundaries. Some European scientists divide the field into :1) Red biotechnology2) Green biotechnology Some divides it into :1) White2) Blue Biotechnology falls under many umbrellaswhich is basically considered as life science.Book : Biotechnology & Genetic engineering (Kathy wilson peacock)52010,Edi:1 : Page No. 4 (Chapter 1)5 6. Biology & ZoologyCell biologyMicrobiologyMolecular BiologyPhysiology, Ecology,EmbryologyGenetics,Population genetics,EpigeneticsProteonomics &BioinformaticsBook : Biotechnology & Genetic engineering66 7. 7INTRODUCTION continuation.. Genetics science of genes, heredityand variation in living organisms. Genetics deals with the molecular structureand function of genes, and gene behavior incontext of a cell or organism (e.g. dominanceand epigenetics ). Patterns of inheritance from parent tooffspring, and gene distribution, variation andchange in populations = Population genetics.7Book : Genetics and the Organism: Introduction 8. 8INTRODUCTION continuation..EssenceOfGenetics Chromosome Packaged and organized chromatin, a complex ofmacromolecules found in cells, consisting of DNA, protein andRNA.EssenceOfGenetics DNA A molecule that encodes the genetic instructions used in thedevelopment and functioning of all known living organisms andmany viruses.EssenceOfGenetics Genetic Variation Permanent change in the chemical structure of genes broughtabout by mutation, important in providing genetic material fornatural selection.EssenceOfGenetics Heredity The study of heredity in biology is called genetics, which includesthe field of epigenetics.8Book : Genetics and the Organism: Introduction 9. 99U S National Library of Medicine 10. 1010A form 11 bp/ turnB form-10 bp/ turnZ form-12 bp/ turnFrom Lippincotts - Illustrated Biochemistry 11. 1111Deletion Inversion DuplicationInsertion Translocation 12. 12 13. INTRODUCTION continuation..Various Branches of GeneticsBehaviouralgeneticsClassicalgeneticsDevelopmentalgeneticsConservationgeneticsEcologicalgenetics EvolutionarygeneticsGeneticengineering&Metagenesis1313Book : Genetics and the Organism: Introduction 14. CONTENTS1414 15. BASICS OFGENETIC ENGINEERING Different terms used for geneticengineering :1) Gene manipulation2) Gene cloning3) Recombinant DNA technology4) Genetic modification5) New geneticsAn Introduction to Genetic Engineering (Desmond S. T. Nicholl) Edi :3rd 200815Chapter 2 . Page 315 16. Direct manipulation of an organism'sgenome using biotechnology .First isolating andcopying thegenetic material ofinterestusing molecularcloning methodsGenerate aDNA sequenceNew DNAinserted in the hostgenomeBASICS OFGENETIC ENGINEERING CONTINUATION..An Introduction to Genetic Engineering (Desmond S. T. Nicholl)Edi :3rd 2008 Chapter 2.16 17. 17Miller et al(2000). An Introduction to Genetic Analysis (7th ed.).17 18. 1818CONTENTS1) INTRODUCTION2) BASICS OF GENETIC ENGINEERING4) GENETIC ENGINEERING IN DENTISTRY- VARIOUS TECHNIQUES OF GENETIC ENGINEERING5)GENETIC COUNSELLING AND ROLE OF A DENTIST1) SUMMARY2) CONCLUSION3) REFERENCES 19. Genetic inheritance was firstdiscovered by Gregor Mendel in 1865following experiments crossing peas. Although largely ignored for 34 years heprovided the first evidence of hereditarysegregation and independent assortmentIn 1889 Hugo de Vries came up with the name"(pan)gene" for after postulating that particlesare responsible for inheritance of characteristicsTerm "genetics" was coined by WilliamBateson in 1905.19 20. In 1928 Frederick Griffith proved theexistence of a "transforming principle" involvedin inheritance, which Avery, MacLeod andMcCarty later (1944) identified as DNA.Edward Lawrie Tatum and George WellsBeadle developed the central dogma that genescode for proteins in 1941.The double helix structure of DNA wasidentified by James Watson and FrancisCrick in 1953.20 21. In 1970 Hamilton Smiths labdiscovered restriction enzymes that allowedDNA to be cut at specific places and separatedout on an electrophoresis gel. This enabled scientists to isolate genes from anorganism's genome.DNA ligases, that join broken DNA together,had been discovered earlier in 1967 and bycombining the two enzymes it was possible to"cut and paste" DNA sequences tocreate recombinant DNA.Plasmids, discovered in 1952, becameimportant tools for transferring informationbetween cells and replicating DNA sequences.21 22. Frederick Sanger developed a method forsequencing DNA in 1977, greatly increasing thegenetic information available to researchersPolymerase chain reaction (PCR), developedby Kary Mullis in 1983, allowed small sectionsof DNA to be amplified and aided identificationand isolation of genetic materialArtificial competence was inducedin Escherichia coli in 1970 when MortonMandel and Akiko Higa showed that it couldtake up bacteriophage after treatment withcalcium chloride solution (CaCl2).22 23. Two years later, Stanley Cohen showed thatCaCl2 treatment was also effective for uptake ofplasmid DNA.Transformation using electroporation wasdeveloped in the late 1980s, increasing theefficiency and bacterial rangeIn 1972 Paul Berg utilised restriction enzymesand DNA ligases to create the first recombinantDNA molecules.23 24. Herbert Boyer and Stanley N. Cohen tookBergs work a step further and introducedrecombinant DNA into an bacterial cell.In 1981 the laboratories of Frank Ruddle,Frank Constantini and Elizabeth Lacyinjected purified DNA into a single-cell mouseembryo and showed transmission of the geneticmaterial to subsequent generations.On June 19, 2013 the leaders of three researchteams who originated the technology, Robert T.Fraley of Monsanto; Marc VanMontagu of GhentUniversity in Belgium and founder of Plant GeneticSystems and CropDesign ; and Mary-DellChilton ofWashington University in St.Louis and Syngenta were awarded with the WorldFood PrizeGordon, J.; Ruddle, F. (1981). "Integration and stable germ line transmission 24ofgenes injected into mouse pronuclei". Science 214 (4526): 1244. 25. The first recorded knockout mouse was createdby Mario R. Capecchi, MartinEvans and Oliver Smithies in 1989. They areused to study gene function and make usefulmodels of human diseases.In 1992 onco-mice with tumor suppressorgenes knocked out were generated.Creating Knockout rats are much harder and hasonly been possible since 2003Bacteria synthesising human insulin weredeveloped in 1979, being used as a treatment forthe first time in 1982Zan, Y; Haag, J. D.; Chen, K. S.; Shepel, L. A.; Wigington, D; Wang, Y. R.; Hu, R; Lopez-Guajardo, C.C.; Brose, H. L.; Porter, K. I.; Leonard, R. A.; Hitt, A. A.; Schommer, S. L.; Elegbede, A. F.; Gould, M.N. (2003). Production of knockout rats using ENU mutagenesis and a yeast-based screening25assay". Nature Biotechnology 21(6): 64551. 26. 26In 1988 the first human antibodies wereproduced in plants.The first animal to synthesise transgenicproteins in their milk were mice, engineeredto produce human tissue plasminogenactivator.With the discovery of microRNA in 1993came the possibility of using RNAinterference to silence an organismsendogenous genes- Peng, S. (2006). "A transgenic approach for RNA interference-basedgenetic screening in mice". Proceedings of the NationalAcademy of Sciences 103 (7): 22522220.- Vaucheret, H.; Chupeau, Y. (2011). plant miRNAs regulate geneexpression in animals Cell Research 22 (1): 35. 27. 27 Improved ourunderstanding ofgenetics. His research helped tomake organtransplantationspossible.Dr. Bernard Amos27 28. His work cloned frogslaid the foundationsfor somatic cellnuclear transfer, theapplication of whichled to Dolly the sheep.28John Gurdon28 29. Worked out theStructure ofProteins.29Linus Pauling29 30. 30The Father of CloningHans Spermann30 31. 31The Father of GeneticsGregor Mendel31 32. He noticed that there isa pattern in the 4 bases:Adenine, Guanine,Cytosine and Thymine.32 A=T and G=C.Erwin Chargaff32 33. In 1973 createda transgenicmouse byintroducing foreignDNA into its embryo,making it the worldsfirst transgenicanimal.33Rudolf Jaenisch33 34. 3434CONTENTS1) INTRODUCTION2) BASICS OF GENETIC ENGINEERING3) HISTORY OF GENETIC ENGINEERING5) GENETIC ENGINEERING IN DENTISTRY- VARIOUS TECHNIQUES OF GENETIC ENGINEERING6) 5)GENETIC COUNSELLING AND ROLE OF A DENTIST7) SUMMARY8) CONCLUSION9) REFERENCES 35. GENERAL APPLICATIONS OFGENETIC ENGINEERING][38]35Eg: transgenic plants produce naturalpesticide to resist to pest35 36. Engineered MammalsA monkey named ANDi,for "inserted DNA", in apicture released inJanuary 2001. ANDi wasborn in October 2000 atthe Oregon HealthScience University afterreceiving an extra bit ofgenetic material tobecome the world's firstgenetically modifiednon-human primate3636 37. 37Cloning Dolly Sheep A: donate body cell nucleus Sheep B: donate an egg cell without nucleus Sheep C: surrogate motherA B CDolly37Whos itsmother? 38. 3838CONTENTS1) INTRODUCTION2) BASICS OF GENETIC ENGINEERING3) HISTORY OF GENETIC ENGINEERING4) GENERAL APPLICATION OF GENETIC ENGINEERING6) 5)GENETIC COUNSELLING AND ROLE OF A DENTIST7) SUMMARY8) CONCLUSION9) REFERENCES 39. GENETICS & ORAL HEALTH3939Craniofacial &ToothmorphogenesisAgenesisGENETICSDental cariesPeriodontistisCleft lip &PalateGeneticdisorders /AbnormalitiesOral Cancer MalocclusionBehavorialGeneticsPharmacogenetics 40. TECHNIQUESOFGENETIC ENGINEERINGTools and techniquesMethods in recombinant DNAtechnologyGenetically modified organismsGenetic treatments4040 41. 41TECHNIQUESOFGENETIC ENGINEERINGMethods in recombinant DNAtechnologyGenetically modified organismsGenetic treatments41 42. DNA: The Raw Material Heat-denatured DNA DNA strands separate if heatedto just below boiling Exposes nucleotides Can be slowly cooled andstrands will renature4242 43. Restriction Endo-nucleases Enzymes that can clip strands of DNAcrosswise at selected positions Each has a known sequence of4 to 10 pairs as its target Can recognize and clip at palindromes4343 44. Can be used to cut DNA in to smallerpieces for further study or to removeand insert sequences. Can make a blunt cut or a sticky end The pieces of DNA produced arecalled restriction fragments. Differences in the cutting pattern of specificrestriction endonucleases give rise torestriction fragments of differing lengths-restrictionfragment length polymorphisms.4444 45. 4545 46. Ligase and Reverse Transcriptase Ligase: Enzyme necessary to sealsticky ends together Reverse transcriptase: enzyme that isused when converting RNA into DNA.4646 47. 4747 48. ANALYSIS OFDNAGelelectrophoresisPolymeraseChain Reaction4848 49. 4949 Gel electrophoresis: produces a readablepattern of DNA fragments 50. GEL ELCTROPHORESIS APPLICATIONS:Estimation of the size of DNAmolecules following restrictionenzyme digestion, e.g. in restrictionmapping of cloned DNA.Analysis of PCR products, e.g. inmolecular genetic diagnosis or geneticfingerprinting.5050 51. 5151 Some techniques to analyze DNA andRNA are limited by the small amounts oftest nucleic acid available Polymerase chain reaction (PCR)rapidly increases the amount of DNA in asample So sensitive- could detect cancer from asingle cell Can replicate a target DNA from a fewcopies to billions in a few hours 52. 5252 53. 5353Three Basic Steps that Cycle Denaturation Heat to 94C to separate in to two strands Cool to between 50C and 65C Priming Primers added in a concentration that favors bindingto the complementary strand of test DNA Prepares the two strands (amplicons) for synthesis Extension 72C DNA polymerase and nucleotides are added Polymerases extend the molecule The amplified DNA can then be analyzed 54. 5454 Relative sizes of nucleic acidsusually denoted by the number ofbase pairs (bp) they contain. DNA Sequencing: Determining theExact Genetic Code Most detailed information comes fromthe actual order and types of bases-DNA sequencing Most common technique: Sanger DNAsequence technique 55. 5555 56. 5656 Two different nucleic acids can hybridize by uniting attheir complementary regions. Gene probes: specially formulated oligonucleotidetracers Short stretch of DNA of a known sequence Will base-pair with a stretch of DNA with acomplementary sequence if one exists in the test sample Can detect specific nucleotide sequences in unknownsamples. Probes carry reporter molecules (such as radioactive orluminescent labels) so they can be visualized. Southern blot- a type of hybridization technique 57. 5757 Southern blotting involves the transfer of DNAfrom a gel to a membrane, followed by detectionof specific sequences by hybridization with alabeled probe. Northern blotting, RNA is run on a gel. Western blotting entails separation of proteins onan SDS gel, transfer to a nitrocellulose membrane,and detection proteins of interest using antibodies. 58. 58 FIGURE 21: Southern blot: Identifying Specific DNA Fragments(Edward Southern--the pioneer)or gentle vacuumpressureDrying or exposureto UV lightProbes: Isotope or chemicalGel is soaked inalkali buffer todenature DNA 59. Northern blotting is similar to Southern blotting,but involves the transfer of RNA from a gel to amembraneRNA59 60. Northern blotting: Measuring gene activityPoly(A)+ RNA: from rat tissuesProbe: G3PDH (glyceraldehyde-3-phosphate dehydrogenase)60 61. Western blotting61 Western blotting entails separation of proteins onan SDS gel, transfer to a nitrocellulose membrane,and detection proteins of interest using antibodies.wikipedia 62. Western blot 62 63. Blotting Methods63 Antibodies can recognize the protein of interest oran epitope tag. epitope tag A short peptide sequence thatencodes a recognition site (epitope) for anantibody, typically fused to a protein of interest fordetection or purification by the antibody.Human influenza hemagglutinin (HA): YPYDVPDYAThe HA tag is derived from the HA-molecule corresponding to aminoacids 98-106 has been extensively used as a general epitope tag inexpression vectors. 64. 6464 65. 6565 Probes applied to intact cells Observed microscopically for thepresence and location of specificgenetic marker sequences Effective way to locate genes onchromosomes 66. Gene expression array are used to detect thelevel of all the expressed genes in anexperimental sample. SNP arrays permit genome-wide genotyping ofsingle nucleotide polymorphisms. =>useallele-specific oligonucledtide probe Array comparative genome hybridization(array-CGH) allows the detection of copynumber changes in any DNA sequencecompared between two samples.66 67. DNA 67Microarrays DNA microarrayscomprise known DNAsequences spotted orsynthesized on a smallchip.Microarrays show thelevels of all theexpressed genes in anexperimental sample. 68. 68TECHNIQUESOFGENETIC ENGINEERINGTools and techniquesGenetically modified organismsGenetic treatments68 69. 6969Methods in Recombinant DNATechnology Primary intent of recombinant DNAtechnology- deliberately removegenetic material from one organism andcombine it with that of a differentorganism. Form genetic clones Gene is selected Excise gene Isolate gene Insert gene into a vector Vector inserts DNA into a cloning host 70. 7070Methods in Recombinant DNATechnology 71. 7171Technical Aspects of RecombinantDNA and Gene Cloning Strategies for obtaining genes in anisolated state DNA removed from cells, separated intofragments, inserted into a vector, andcloned; then undergo Southern blotting andprobed Gene can be synthesized from isolatedmRNA transcripts Gene can be amplified using PCR Once isolated, genes can be maintainedin a cloning host and vector (genomiclibrary) 72. 7272Characteristics of CloningVectors Capable of carrying a significant piece of thedonor DNA Readily accepted by the cloning host Must have a promoter in front of the clonedgene Vectors (such as plasmids andbacteriophages) should have three importantattributes: An origin of replication somewhere on thevector Must accept DNA of the desired size Contain a gene that confers drug resistance totheir cloning host 73. 7373 74. 7474Characteristics of Cloning Hosts 75. 75APPLICATIONSOFGENETIC ENGINEERINGTools and techniquesMethods in recombinant DNAtechnologyGenetically modified organisms75 76. 7676TREATMENT OFGENETIC DISEASE Conventional approach Gene Therapy 77. CONVENTIONAL APPROACH7777OF TREATMENT Enzyme induction by drugs Replacement of deficient enzymes /proteins Replacement of deficient vitamin / co-enzyme Replacement of deficient product Substrate restriction in diet Drug therapy Drug avoidance Replacement of diseased tissue Removal of disease tissue 78. Genomic medicine 78use of genotypic analysis (DNAtesting) to enhance quality of medicalcare, including78- presymptomaticidentification- preventive intervention- selection ofpharmacotherapy- design of medical care 79. 7979GENE THERAPYReplacement of a deficient gene /gene product or correction of anabnormal gene.2 TYPES:i. Germ-line gene therapy changeswill be passed on to subsequentgenerationsii. Somatic Cell gene therapy changes will not be passed on tofuture generations 80. Gene Therapy Gene transfer for the purpose oftreating human disease. Transfer of new genetic material aswell as manipulation of existinggenetic material.(Genetic engineering)in vivo ex vivo80 81. 81 82. Gene therapyPotential Uses Treatment of recurrent disease Adjuvant treatment Localized distant metastatic disease82 83. Delivery systems / vectors Non viral- Electropolation- DEAE-dextran- Calcium phosphate- Liposomes-Naked DNA VirusesRetrovirusesAdenovirusesAdeno-associated virusesHerpes virusGene therapy83 84. Gene therapy84 85. Gene therapy in dentistry1. Bone repair Mesenchymal stem cell mediatedgene therapy (BMPs) Using adenoviral vector Transfer of Platelet derivative growthfactor (PDGF) Bone sialoprotein delivery (in-vivo)2. Salivary glands Irreversible salivary glanddysfunctionGene therapy85 86. Gene therapy Adenovirus encoding human AQP1(water channel protein) irradiatedsalivary gland hyposalivation. Autoimmune diseasesSjogren syndrome :cytokines inflammationadeno-associated virus, AAV,serotype2 IL-10 transfer usingrecombinant AAV2 vector salivary glands hyposalivation .86 87. Gene therapy8787 88. Gene therapeuticsGene therapy88local (exocrine) systemic (endocrine)(oral, pharyngeal, (single protein &esophageal) deficiency)Eg mucosal cadidiasis Eg hGH Azole resistant Recombinant adenoviral vector encoding H3 89. Gene therapy PainVirus vector mediated transfer of genesencoding opiate peptidesperipheral & central neuronsAnti-noceptive effects89Direct gene delivery articular surface TMJ 90. Keratinocytes Gene therapy systemichuman aplipoprotein E, factor IX,growth hormone and IL-10 intobloodstream. DNA vaccinationsGene therapy90 91. Gene therapy Gene gun-based DNA vaccination againstinfectious diseases oral mucosa(Wang J 2003) Caries vaccine91 92. Subunit VaccinesGene therapy- synthetic peptide vaccines- recombinant vaccines Conjugate Vaccines Routes to Protective Responses- oral- intranasal- tonsillar- rectal92 Adjuvants and Delivery SystemsCholera & E coli, microcapsules, liposomes 93. Human applicationsGene therapy93- Active immunization ( 7 trials)- Passive immunization ( cows milk,chicken eggs, transgenic plant antibody) 94. Gene therapy94Future Strategies of Gene Therapyin Preventing Periodontal Diseases Gene Therapeutics-PeriodontalVaccination Genetic Approach to BiofilmAntibiotic Resistance An In vivo Gene Transfer byElectroporation for AlveolarRemodelling Tight Adherence Gene for the Controlof Periodontal Disease Progression Antimicrobial Gene Therapy toControl Disease Progression 94 95. Gene therapy9595 96. AIDS vaccine HPV vaccine HSV vaccine Head & neck cancerGene therapy96 97. Current gene therapies for cancer 97Mechanism Goal DevelopmentstageOncogenedown-regulationtherapyDeletedefectivegeneInhibit tumorcell growthPre-clinicalGeneadditiontherapyAdd tumorsuppressorgeneKill tumor cell Clinical trialAnti-senseRNAAbrogategenesstimulatingtumor growthInhibit tumorcell growthClinical trialImmunotherapyEnhanceimmunesurveillanceEnhanceimmunogenicity of tumorcellClinical trial 98. Anti-angiogenesistherapyTransfergene totumor cellsto blockangiogenesisInhibit tumorprogressionPre-clinicalDrugresistancegene therapyTransfercytoprotective geneDecreasetoxicity ofchemotherapyClinical trialTumor-cellkillingvirusesIntroduceviruses thatdestroytumor cellsas part ofreplicationcycleKill tumorcellsPre-clinicalSuicide genetherapyTransfergeneencodingpro-drugactivatingenzymeKill tumorcell &enhancechemotherapyClinical trial98 99. 99 HUMAN GENOME99PROJECTObjectives:i. Sequencing of human genomesii. Mapping of human inherited diseasesiii. Development of new DNAtechnologiesiv. Development of bio-informaticsv. Comparitive Genomicsvi. Functional Genomics 100. 100TISSUE ENGINEERING Tissue Engineering is a general nameof biomedical fields to enable cells toenhance their proliferation,differentiation, and morphologicalorganization for induction of tissueregeneration, resulting in regenerativemedical therapy of diseases.100 101. 101Stem cells in regenerative medicine A stem cell is defined as a cell that cancontinuously produce unaltereddaughters and, furthermore, has theability to generate cells with differentand more restricted properties. These cells can either multiply(progenitors or transit amplifying cells)or be committed to terminaldifferentiation. Stem cells are self-renewing and thuscan generate any tissue for a lifetime. This is a key property for a successfultherapy. 101 102. 102102 103. 103103 104. 104104 105. 105105 106. 106106 107. 107GENETIC COUNSELLINGA process of communication and educationwhich addresses concerns relating to thedevelopment and / or transmission of ahereditary disorder.STEPS IN GENETIC COUNSELLING107- Diagnosis- Risk assessment- Communication- Discussion of options- Long term contact & support 108. 108108DIAGNOSIS History Examination Investigation Only when accurate diagnosis ispossible When etiological heterogeneity ispresent 109. 109RISK ASSESSMENTThe good side of the coin should alsobe emphasized109ARBITRARY GUIDE 1 in 10 - High risk 1 in 20 - Low risk Intermediate values - Moderate risk 110. 110LONG TERM CONTACT &110SUPPORT Counselling centers should maintaininformal contact with familiesthrough a network of geneticassociates Genetics registers provide a usefulmeans in ensuring effective contact 111. 111NEONATAL SCREENING111To prevent subsequent morbidityPOPULATION CARRIERSCREENINGThe branch of medical geneticswhich is concerned withscreening and the prevention ofgenetic disease on a populationbasis is known as communitygenetics. 112. PRENATAL DIAGNOSIS 112Ability to detect abnormality in anunborn child.112TECHNIQUESI. Non invasive- Maternal Serum screening- Ultra soundII. Invasive- Amniocentesis- Chorionic Villus Sampling 113. INDICATIONS FOR 113113PRENATAL DIAGNOSIS Advanced maternal age Previous child with a geneticabnormality Family History of- Chromosome abnormality- Single gene disorder- Neural tube defect- Other congenital structuralabnormalities Abnormalities identified in pregnancyEg. Poor fetal growth 114. 114114 Other High risk factors- Parental Consanguinity- Poor obstetric historyEg: Recurrent miscarriagesPrevious unexplained stillbirth- Maternal illnessEg: Poorly controlled IDDMMaternal epilepsyTreatment with SodiumValproate 115. IDENTIFY GENETIC DISEASE1. Build the pedigree2. Analyse3. Risk of recurrence4. Decision115 116. Role of dentist as geneticcounselor Oral manifestations Correct identification Diagnosis Referral Suggestion Screening for dental diseasesDNA probes116 117. Future prospects117 Bioengineering Nanodentistry Biomimetics Molecular Epidemiology ( VariationGenetics ) 118. 118118 119. 119GeneticengineeringEnablingtechnologyCutting,modifyingand joining DNAmoleculesenzymesGenerationof DNAfragmentsRestrictionenzymeDNA LigaseJoining to avector or DNAMoleculeIntroductioninto the hostcellSelection ofdesiredsequenceArose fromGene cloningRecombinantDNAMolecular cloningPure science,Biotechnology,Medicine,DentistryLegal andethicalconsiderationsMicrobial &MoleculargeneticsIn 1972StanfordUniversityIs also known asHas application inBut raises somewas first achievedIs anThat involvesusingSuch asRequires four stepsCan be used forused for 120. CONCLUSION120 Biotechnology as a fast developingtechnology as well as science , hasalready shown its impact on differentaspects of day-to-day human life suchas public health pharmaceuticals, foodand agriculture industries,bioenergetics and informationtechnology.120 121. 121 As it has potential to ensure foodsecurity, dramatically reduce hungerand malnutrition and reduce ruralpoverty , particularly in developingcountries , Now it is very clear thatbiotechnology is the key technologyfor the 21st century and the science ofthe future.121 122. References1. Colin EC. What Is Genetics? In: Colin EC. Elements of Genetics.3rd ed. New York: McGraw Hill Book Company, Inc; 1956. p. 1-3.2. Auden WH. History of Human Genetics. In: Motulsky V. HumanGenetics, Problems and Approaches. 3rd ed. New York: Springer;1997. p. 1-22.3. Dhar PK. Genetics in pediatric dentistry. In: Tandon S. Textbook ofPedodontics. Ist ed. Hyderabad: Paras Medical Publisher; 2001. p.614-622.4. Baeudet AL. Genetics and Disease. In: Fauci, Braunwald,Isselbacher, Wilson, Martin, Hauser, Longo. Harrisons Principlesof Internal Medicine. Vol 2. 14th ed. New York: McGraw HillCompanies, Inc; 1998. p. 365-395. 123. 5 Tortora GJ, Grabowski SR. Cellular level of organization. 10th ed. Johnwiley & sons, USA. 84-97.6 Pashayan HM, Feingold M. Selected syndromes in pedodontics.White GE. Clinical oral pediatrics. Quintessence International. Tokyo.1981. ed. 73-80.7 Joerde LB, Carey JC, White RL. Genetic Variation: Its Origin andDetection. In: Joerde LB, Carey JC, White RL. Medical Genetics. Isted. Toronto: Mosby; 1995. p. 30-56.8 Tencate AR et al. Development of tooth and its supporting structures.In: Nanci A. Tencates oral histology. 8thed. Mosby. 2003: 79-110.9 Tandon S, Bhat S. Developing dentition and its disturbances. In:Tandon s. Textbook of pedodontics. Ed. Paras publishing . 2001. 85-105. 124. 10. Shafer. Textbook of oral pathology. 4th ed. Harcourt Asia PTELtd.. 1999.11 Anderson M. Risk Assessment and Epidemiology of DentalCaries: Review of Literature. Pediatr Dent 2002; 24(5): 377-385.12 Sofaer JA. Host genes and dental caries. Br Dent J 1993; 175:403-409.13 Shuler Cf. Inherited risks for susceptibility to dental caries. JDent Educ 2001; 65(10): 1038-1045.14. Hart TC & Kornman KC. Genetic factors in pathogenesis ofperiodontitis. Periodont 2000 1997; 14: 202-15.15 Michalowicz. Genetic & heritable risk factors in periodontaldisease. J Periodontol 1994; 65: 479-88. 125. 16 Cobourne MT. the complex genetics of cleft lip & palate. Euro JOrtho 2004; 26: 7-16.17 Mossey PA et al. The heritability of malocclusion. The influence ofgenetics in malocclusion. Br J Ortho 1999; 26: 195-203.18 Partridge M. Oral cancer: the genetic basis of disease. Dent Update2000; 27: 242-248.19 Carrozzo M. Hepatitis C virus-associated oral lichen planus: isthe geographical heterogeneity related to HLA-DR6?J OralPathol Med. 2005 Apr;34(4):204-8.20 Lawson CAet al. Analysis of the insertion/deletion relatedpolymorphism within T cell antigen receptor beta variable genesin primary Sjogren's syndrome.Ann Rheum Dis. 2005Mar;64(3):468-70. 126. 24 Campisi G. HPV DNA in clinically different variants of oralleukoplakia and lichen planus. Oral Surg Oral Med OralPathol Oral Radiol Endod. 2004 Dec;98(6):705-11.25 Xi S etal. Gene therapy for the treatment of oral squamous cellcarcinoma. J Dent Res 2003; 82: 11-16.26 Baum JB. The impact of gene therapy on dentistry. JADA2002; 133: 35-44.27 Wang J et al. Predominant cell-mediated immunity in the oralmucosa: gene gun-based vaccination against infectiousdiseases. J Dermatol Sci. 2003 May;31(3):203-10.28 Slavkin HC. And the next 50 years. The future of DNAtechnology in oral medicine. JPHD 1996; 56: 278-85. 127. 29. [email protected] . [email protected] .Faiez N. Genetics. DCNA. 1975. 1-150.36. Wise GE. Cell and molecular biology of tooth eruption. In:Davidowitch Z. Biologic mechanisms of tooth eruption. Ed.Harvard society. Boston. 1998. 1-7. 128. 37. Griffiths, Anthony J. F.; Miller, Jeffrey H.; Suzuki, David T.;Lewontin, Richard C.; Gelbart, William M., eds. (2000). "Geneticsand the Organism: Introduction". An Introduction to GeneticAnalysis (7th ed.). New York: W. H. Freeman.38. Book : Biotechnology & Genetic engineering (Kathy wilsonpeacock) 2010,Edi:1 : Page No. 4 (Chapter 1)39. D. L. Hartl and V. Orel (1992). "What Did Gregor Mendel ThinkHe Discovered?".Genetics 131 (2): 2452540. Zambryski, P.; Joos, H.; Genetello, C.; Leemans, J.; Montagu, M.V.; Schell, J. (1983). "Ti plasmid vector for the introduction of DNAinto plant cells without alteration of their normal regenerationcapacity". The EMBO Journal 2 (12): 21432150.128