Mis-sense mutationNon-sense mutation

Large deletionConsensus splice sites mutation

Promoter defects

Identification of possible disease causing mutations by

systematic genomic sequencing of

candidate genes

Functional genomics and pre mRNA

processing alterations

Polymorphisms, atypical and “orphan” mutations

Functional splicing assay

Splicing defects

Clear possible functional

alteration

Sal I BamH1

polyA tail promoter

polyA tail

Fibronectin-Globin hybrid

minigene

SXN13 hybridminigene

promoter

promoterpolyA tail

aa

TCGACGTTNNNNNNNNNNNNNGAATG GCANNNNNNNNNNNNNNCTTACCTAG

Can you make the drawings as linear constructs?

aa

TCGACGTTNNNNNNNNNNNNNGAATG GCANNNNNNNNNNNNNNCTTACCTAG

Mis-sense mutationNon-sense mutation

Large deletionConsensus splice sites mutation

Promoter defects

Identification of possible disease causing mutations by systematic genomic sequencing of candidate genes

Analysis of pre mRNA processing alterations

Polymorphisms, atypical and “orphan” mutations

Functional splicing assay (reporter genes)

Identification of mutations as possible Splicing defects

Clear possible functionalalteration

Identification of candidate genes by clinical diagnosis

RNA expression analysis

Verification that mutations act on splicing

Identification of trans-acting factorsDetermination of disease causing mechanism

Development of rational therapy approaches, evidence based genetic conseling

Clin

ical

evalu

ation

B

ioin

form

atic ana

lysis

Exp

erimen

tal valid

ation