Dr. Derakhshandeh, PhD Mutation Screening. 2 TYPE OF MUTATIONS WHICH TECHNIQUES DETECT WHAT TYPE OF...

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Dr. Derakhshandeh, PhD Mutation Screening Mutation Screening

Transcript of Dr. Derakhshandeh, PhD Mutation Screening. 2 TYPE OF MUTATIONS WHICH TECHNIQUES DETECT WHAT TYPE OF...

Page 1: Dr. Derakhshandeh, PhD Mutation Screening. 2 TYPE OF MUTATIONS WHICH TECHNIQUES DETECT WHAT TYPE OF MUTATIONS In classical genetics, three types of mutations.

Dr. Derakhshandeh, PhD

Mutation ScreeningMutation Screening

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TYPE OF MUTATIONSTYPE OF MUTATIONS

WHICH TECHNIQUES DETECT WHAT TYPE OF MUTATIONS

In classical genetics, three types of mutations are distinguished:

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Different types of Different types of mutationsmutations

genome mutations: changes in chromosome number

chromosome mutations: changes in chromosome structure

gene or point mutations: mutations where changes are at molecular level

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genome genome mutations:mutations:

changes in changes in chromosome chromosome

numbernumber

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TechniquesTechniques

Karyotyping, conventional cytogenetics

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Down Syndrome Down Syndrome ((Trisomy 21(Trisomy 21(

Trisomy 2(Trisomy 2(

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chromosome mutations: chromosome mutations: changes in chromosome changes in chromosome

structurestructure

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CHANGES IN CHROMOSOME CHANGES IN CHROMOSOME STRUCTURESTRUCTURE

TranslocationsLarge Deletions/InsertionsInversionsDuplications/Amplifications

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TechniquesTechniques

Conventional cytogenetics molecular cytogenetics

FISHMolecular:

PFGE, Southern blotting, Northern BlottingFluorescence Dosage analysis

large deletionsInsertionsduplications

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Interphase FISH Examples

13 (green),13 (green), and 21 (red) and 21 (red)18 (aqua),18 (aqua), X (green),X (green), and Y (red). and Y (red).

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gene or point gene or point mutations:mutations:

mutations where mutations where changes are at changes are at molecular levelmolecular level

Page 12: Dr. Derakhshandeh, PhD Mutation Screening. 2 TYPE OF MUTATIONS WHICH TECHNIQUES DETECT WHAT TYPE OF MUTATIONS In classical genetics, three types of mutations.

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Methods for Methods for detection of known detection of known

mutationsmutations

Methods for Methods for detection of unknown detection of unknown

mutationsmutations

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Methods for detection of Methods for detection of known mutationsknown mutations

PCR and size separation eg. DMD PCR and restriction enzyme digestion eg. SMN

exon 7 & 8 Allele specific amplification (ASA)

Allele refactory mutation system (ARMS) eg. CF

Allele specific oligonucleotide hybridisation (ASO) Dot Blot eg. CF DNA chips eg. Brca1

Genomic DNA sequencing

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ARMSARMS

Amplification Refractory Amplification Refractory Mutation System Mutation System

Allele Specific PCR (ASPCR) PCR Amplification of Specific Alleles (PASA)

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UsesPopulation screening

rapid (1 working day) inexpensive non-isotopic

Used for testing for B-thalassaemia Cystic Fibrosis Sickle-cell anaemia Phenylketonuria Apolipoprotein E, etc

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Modifications/Adaptions to the original ARMS methodology

Multiplex ARMS

Page 17: Dr. Derakhshandeh, PhD Mutation Screening. 2 TYPE OF MUTATIONS WHICH TECHNIQUES DETECT WHAT TYPE OF MUTATIONS In classical genetics, three types of mutations.

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To set up multiplex ARMS

Determine commonest mutations in the population

develop the muliplex ARMS these mutations

validate the results of the multiplex test on samples with another methology

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Agarose gels showing the Agarose gels showing the feasibility of the ARMS feasibility of the ARMS conceptconcept

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ARMS-PCRARMS-PCR

1 2 3 4 5 6 7 8 9 10 11 12 N M N M N M N M N M N M

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Methods for detection Methods for detection of unknown of unknown mutationsmutations

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Methods for unknown mutations (diagnostic methods)

These methods are relatively simple, but still require:

experience and skill to perform.

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BRCABRCA

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BRCA1 Gene

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BRCA2 Gene

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SSCP single strand conformation

polymorphism

simplicity clearly by heteroduplex analysis (HA)

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Pedigree of a selected family with breast cancer

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SSCP AnalysisBRCA1 Exon 15, 4650delCA

Page 28: Dr. Derakhshandeh, PhD Mutation Screening. 2 TYPE OF MUTATIONS WHICH TECHNIQUES DETECT WHAT TYPE OF MUTATIONS In classical genetics, three types of mutations.

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Pedigree of a selected family with breast cancer

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SSCP AnalysisBRCA1, Exon 20,Nt 5382

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SSCP AnalysisExon 11pi BRCA1 MS R1347G

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A woman having amniocentesis


Molecular pathology: Physiopathology effect of Mutations Dr Pupak Derakhshandeh, PhD Ass Prof of Medical Science of Tehran University.

Molecular pathology: Physiopathology effect of Mutations Dr Pupak Derakhshandeh, PhD Ass Prof of Medical Science of Tehran University.

The Human Genome and Human Evolution Y Chromosome Dr Derakhshandeh, PhD.

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Dr. Derakhshandeh, PhD Mutation Screening 2 Quantitative PCR and Dosage.

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Mutations in SERPINF1 Cause Osteogenesis Imperfecta Type VI

Mutations in SERPINF1 Cause Osteogenesis Imperfecta Type VI

Human Genetic Mutations. 2 Main Types of Mutations 1.) Chromosomal Mutations 2.) Gene Mutations.

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PharmacogeneticsPharmacogenetics Dr, P Derakhshandeh, PhD Dr, P Derakhshandeh, PhD.

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1 ChromosomesI Dr Pupak Derakhshandeh, PhD Ass Prof Medical Science of Tehran University Email: derakhshandeh@tums.ac.irderakhshandeh@tums.ac.ir Website:

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Canine genetically inherited eye disease mutations...Canine genetically inherited eye disease mutations Gene Mutation annotation Mutation type Disease designation AHT offers DNA test

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1 ChromosomesII Dr Pupak Derakhshandeh, PhD Ass Prof Medical Science of Tehran University Email: derakhshandeh@tums.ac.irderakhshandeh@tums.ac.ir Website:

1 ChromosomesII Dr Pupak Derakhshandeh, PhD Ass Prof Medical Science of Tehran University Email: [email protected]@tums.ac.ir Website:

1 Chromosomes Dr Pupak Derakhshandeh, PhD Ass Prof Medical Science of Tehran University.

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Models of Dps wild type and mutant with 10 designed hydrophobic mutations per subunit.

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6.7 Mutations Mutations Notes.pdf · Gene Mutations – Point Mutations • Frameshift Mutations oBase-pair insertion or deletion o The result is frequently a nonfunctional protein.

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Mutations in RAS pathway genes correlate with Type of ...

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S Deficiency Type I: Identification of Point Mutations in

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Type IV procollagen missense mutations associated with defects

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Hypertensions pulmonaires pédiatriquesIdentification of PAH genes mutations in a pediatric population Results • No mutations in children with group 3 and type 4 APAH-CHD. • 8

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Description of two new ABCB11 mutations responsible for ...downloads.hindawi.com/journals/cjgh/2011/534918.pdf · of two new ABCB11 mutations responsible for type 2 benign recurrent

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Characterization of NF1 frameshift mutations in pediatric ...Neurofibromatosis type I is an autosomal dominant disease with complete penetrance, caused by mutations in neurofibromin

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