MarwanAlhalabiMDPhDProfessorinReproductiveMedicineFacultyofMedicineDamascusUniversity

&ClinicalMedicalDirectorOrientHospitalAssistedReproductionCenterDamascus,Syria.

• The Human GenomeProject goals :

• To determine thenucleotide sequence allDNA in the humangenome.

• To identify the locationand sequence of everyhuman gene.

• Genome: AlloftheDNAforanorganism

• HumanGenome• Nucleus:3billionbasepairspackagedintochromosomes

• Mitochondrion:16,600basepairspackagedinonecircularchromosome

nGenes=specificsequencesonDNAthatcanbeexpressedintoproteinsandothermolecules.

nGenomes=allDNAinnucleusofthecell.

1. Transcriptional control sequences:bind transcription factors that can activate or inhibittranscription

2. Promoter: recruits RNA polymerase and marks thestart of the transcribed region

3. Transcript: corresponds to the RNA sequenceCan be further subdivided into:

4. Termination sequences: required to terminatetranscription

• DNA copy itself:• Replication

• DNA synthesize RNA• Transcription

• RNA synthesize protein• Translation

Two Main Process: Transcription and Translation

DNA molecule

Gene 1

Gene 2

Gene 3

DNA strand

TRANSCRIPTION

RNA

Polypeptide

TRANSLATIONCodon

Amino acid

Central Dogma of Biology� How does the information flow in biological systems?

The central dogma of biology is that information stored in DNA

is transferred to RNA molecules during transcription and to proteins during translation.

Work to keep the cell alive

DNARNA Protein

Transcription Translation

Carries the directions to the cytoplasm

Directions to make proteins are safely stored in the nucleus

• Anychangeingeneticmaterial.

• DNALevel

• ChromosomalLevel

• GenomeLevel

• change in a DNA sequence that affectsgenetic information.

mRNAAmino acids

Mutant DNA

mRNA

DNA

• 1)Duplications

• 2)Deletions

• 3)Translocations

• 4)Inversions

• Theseresultinploidy changes.

• Aneuploidy.

• Plusorminusoneorafewchromosomes.

3n or more = polyploidy

n = haploid (gametes)2n = diploid (normal individual)

• improvementsinmedicine.

• microbialgenomeresearchforfuelandenvironmentalcleanup.

• DNAforensics.

• improvedagricultureandlivestock.

• betterunderstandingofevolutionandhumanmigration.

• moreaccurateriskassessment.

• Improved diagnosis of diseasesand help with early detectionof genetic disorders

• GeneTherapy.

• Swappinganabnormalgeneforanormalone.

• Repairinganabnormalgene

• improved agriculture :• Help create disease, insect, drought

resistant, and more nutritious plants.

• Scientists transferred daffodilgenes into rice.

• Rice with beta-carotene mayhelp prevent vitamin Adeficiencies.

GoldenRice- geneticallymodifiedricewithVit A.

GeneticallyModifiedOrganisms(GMOs)

•Studyevolution throughgermline mutationsinvariouslineages.

•Studymigration ofdifferentpopulationgroupsbasedonfemalegeneticinheritance.

•StudymutationsontheYchromosometotracelineageandmigrationofmales.

•Greater accuracy in identifying the suspect of a crime by matching their DNA profile with that of any body tissue found at the scene of the crime.

•Paternity testing and other family relationships.

•Exoneration of individual falsely accused of a crime.

•Determine pedigree for seed or livestock breeds

•Assess health damage and risks caused by radiation exposure, includinglow-dose exposures .

•Assess health damage and risks caused by exposure to mutagenicchemicals and cancer-causing toxins.

•Reduce the likelihood ofheritable mutations.

AninternationalefforttodeciphertheDNAblueprintofahumanbeing

• Plannedin1988.• Begunformallyin1990.• Originallyplannedtolast15

years,butrapidtechnologicaladvancesacceleratedcompletiondateto2003.• Announcedè 97%finishedin2000

• FinalHGPpaperswerepublishedin2006.

• Human genome has alot of “junk” so what isthe value of sequencingit?

• Is it too expensive?• HGP cost ~ $3 billiondollars. About $1 perbase pair.

• Fairnessintheuseofgeneticinformation• Whoshouldhaveaccesstopersonalgeneticinformation,andhowwillitbeused?

• Employers• Insurers• Schools• Courts• Adoptionagencies• Military

Genomesinahuman1

Cellsinahumanbody100trillion

Chromosomesinahumancell46

Genesinahumangenome20,500

0

20,000

40,000

60,000

80,000

100,000

120,000

140,000

Early estimates Later estimates Draft sequence Final sequence

More than 99.9% ofhuman DNA sequencesare the same across thepopulation of allhumans in the world.

caggcggactcagtggatctggccagctgtgacttgacaagcaggcggactcagtggatctagccagctgtgacttgacaag

CACACTTGCATGTGAGAGCTTCTAATATCTAAATTAATGTTGAATCATTATTCAGAAACAGAGAGCTAACTGTTATCCCATCCTGACTTTATTCTTTATG AGAAAAATACAGTGATTCC

AAGTTACCAAGTTAGTGCTGCTTGCTTTATAAATGAAGTAATATTTTAAAAGTTGTGCATAAGTTAAAATTCAGAAATAAAACTTCATCCTAAAACTCTGTGTGTTGCTTTAAATAATC

AGAGCATCTGC TACTTAATTTTTTGTGTGTGGGTGCACAATAGATGTTTAATGAGATCCTGTCATCTGTCTGCTTTTTTATTGTAAAACAGGAGGGGTTTTAATACTGGAGGAACAACTGATGTACCTCTGAAAAGAGA AGAGATTAGTTATTAATTGAATTGAGGGTTGTCTTGTCTTAGTAGCTTTTATTCTCTAGGTACTATTTGATTATGATTGTGAAAATAGAATTTATCCCTCATTAAATGTAAAATCAACAGGAGAATAGCAAAAACTTATGAGATAGATGAACGTTGTGTGAGTGGCATGGTTTAATTTGTTTGGAAGAAGCACTTGCCCCAGAAGATACACA

ATGAAATTCATGTTATTGAGTAGAGTAGTAATACAGTGTGTTCCCTTGTGAAGTTCATAACCAAGAATTTTAGTAGTGGATAGGTAGGCTGAATAACTGACTTCCTATC ATTTTCAGGTTCTGCGTTTGATTTTTTTTACATATTAATTTCTTTGATCCACATTAAGCTCAGTTATGTATTTCCATTTTATAAATGAAAAAAAATAGGCACTTGCAAATGTCAGATCACTTGCCTGTGGTCATTCGGGTAGAGATTTGTGGAGCTAAGTTGGTCTTAATCAAATGTCAAGCTTTTTTTTTTCTTATAAAATATAGGTTTTAATATGAGTTTTAAAATAAAATTAATTAGAAAAAGGCA

AATTACTCAATATATATAAGGTATTGCATTTGTAATAGGTAGGTATTTCATTTTCTAGTTATGGTGGGATATTATTCAGACTATAATTCCCAATGAAAAAACTTTAAAAAATGCTAGTG

ATTGCACACTTAAAACACCTTTTAAAAAGCATTGAGAGCTTATAAAATTTTAATGAGTGATAAAACCAAATTTGAAGAGAAAAGAAGAACCCAGAGAGGTAAGGATATAACCTTAC

CAGTTGCAATTTGCCGATCTCTACAAATATTAATATTTATTTTGACAGTTTCAGGGTGAATGAGAAAGAAACCAAAACCCAAGACTAGCATATGTTGTCTTCTTAAGGAGCCCTCCCC

TAAAAGATTGAGATGACCAAATCTTATACTCTCAGCATAAGGTGAACCAGACAGACCTAAAGCAGTGGTAGCTTGGATCCACTACTTGGGTTTGTGTGTGGCGTGACTCAGGTAATC

TCAAGAATTGAACATTTTTTTAAGGTGGTCCTACTCATACACTGCCCAGGTATTAGGGAGAAGCAAATCTGAATGCTTTATAAAAATACCCTAAAGCTAAATCTTACAATATTCTCAA

GAACACAGTGAA ACAAGGCAAAATAAGTTAAAATCAACAAAAACAACATGAAACATAATTAGACACACAAAGACTTCAAACATTGGAAAATACCAGAGAAAGATAAT AAATATTTTACTCTTTAAAAATTTAGTTAAAAGCTTAAACTAATTGTAGAGAAAA

AACTATGTTAGTATTATATTGTAGATGAAATAAGCAAAACATTTAAAATACAAATGTGATTACTTAAATTAAATATAATAGATAATTTACCACCAGATTAGATACCATTGAAGGAATAATTAATATACTGAAATACAGGTCAGTAGAATTTTTTTCAATTCAGCATGGAGATGTAAAAAATGAAA

ATTAATGCAAAAAATAAGGGCACAAAAAGAAATGAGTAATTTTGATCAGAAATGTATTAAAATTAATAAACTGGAAATTTGACATTTAAAAAAAGCATTGTCATCCAAGTAGATGT

GTCTATTAAATAGTTGTTCTCATATCCAGTAATGTAATTATTATTCCCTCTCATGCAGTTCAGATTCTGGGGTAATCTTTAGACATCAGTTTTGTCTTTTATATTATTTATTCTGTTTACTA

CATTTTATTTTGCTAATGATATTTTTAATTTCTGACATTCTGGAGTATTGCTTGTAAAAGGTATTTTTAAAAATACTTTATGGTTATTTTTGTGATTCCTATTCCTCTATGGACACCAAGGC

TATTGACATTTTCTTTGGTTTCTTCTGTTACTTCTATTTTCTTAGTGTTTATATCATTTCATAGATAGGATATTCTTTATTTTTTATTTTTATTTAAATATTTGGTGATTCTTGGTTTTCTCAGC

CATCTATTGTCAAGTGTTCTTATTAAGCATTATTATTAAATAAAGATTATTTCCTCTAATCACATGAGAATCTTTATTTCCCCCAAGTAATTGAAAATTGCAATGCCATGCTGCCATGTG

GTACAGCATGGGTTTGGGCTTGCTTTCTTCTTTTTTTTTTAACTTTTATTTTAGGTTTGGGAGTACCTGTGAAAGTTTGTTATATAGGTAAACTCGTGTCACCAGGGTTTGTTGTACAGATC

ATTTTGTCACCTAGGTACCAAGTACTCAACAATTATTTTTCCTGCTCCTCTGTCTCCTGTCACCCTCCACTCTCAAGTAGACTCCGGTGTCTGCTGTTCCATTCTTTGTGTCCATGTGTTCT

CATAATTTAGTTCCCCACTTGTAAGTGAGAACATGCAGTATTTTCTAGTATTTGGTTTTTTGTTCCTGTGTTAATTTGCCCAGTATAATAGCCTCCAGCTCCATCCATGTTACTGCAAAGA

ACATGATCTCATTCTTTTTTATAGCTCCATGGTGTCTATATACCACATTTTCTTTATCTAAACTCTTATTGATGAGCATTGAGGTGGATTCTATGTCTTTGCTATTGTGCATATTGCTGCAA

GAACATTTGTGTGCATGTGTCTTTATGGTAGAATGATATATTTTCTTCTGGGTATATATGCAGTAATGCGATTGCTGGTTGGAATGGTAGTTCTGCTTTTATCTCTTTGAGGAATTGCCAT

GCTGCTTTCCACAATAGTTGAACTAACTTACACTCCCACTAACAGTGTGTAAGTGTTTCCTTTTCTCCACAACCTGCCAGCATCTGTTATTTTTTGACATTTTAATAGTAGCCATTTTAAC

T

5000basesperpage

• Single-nucleotidepolymorphism• One-nucleotidedifferenceinsequenceoftwoorganisms

• Foundbysequencing

• Example:Betweenanytwohumans,onaverageoneSNPevery1,000basepairs

ATCGATTGCCATGACATCGATGGCCATGAC

• The human genome is nearly the same inall people (99.9%).

• Only 2% of the genome contains genes.• Humans have an estimated 20,500 genes,half of which are still unknown.

• Half of all human proteins sharesimilarities with those of other organisms.

8585

Embryo biopsyDiagnosis

by

Transfer1-2 unaffectedembryos

Fertilisation in vitro (IVF or ICSI)

PCRFISH

Accurate genetic

diagnosisAppropriate Genetic

Counselling

DENATURING

ANNEALING

EXTENSION

PRIMER

TAQ

TAQ

Ovarian Stimulation IVF Blastomere Biopsy on Day 3

Genetic AnalysisTransfer of Unaffected

EmbryoOutcome

Chromosomally Normal Baby

• PolarBody

• CleavageStage

• Blastocyst

Donotprovidediagnosisofpaternalalleles&the

gender theembryo.

• PolymeraseChainReaction(PCR).

• FluorescenceInSituHybridization(FISH).

• CGH24.

• DNAMicroarrays.