TABLE OF CONTENTS
Abstract………..…..i
1. INTRODUCTION………..1
1.1 Genetic causes of isolated CHD……….……...3
1.1-1 Transcription factors and CHD………...4
1.1-2 Somatic mutations………..….9
1.1-3 Novel genetic mutations………...10
1.2 Epigenetics and CHD………....…11
1.2-1 microRNAs……….….12
2. AIMS OF THE STUDY………...………15
3. MATERIAL AND METHODS………16
3.1. Study populations……….……16
3.2. Mutational sequencing analysis……….……17
3.3 In silico analysis for miRNA ≡ SNP interactions……….…..19
3.4 Plasmid construction……….…19
3.5 Cell transfection and luciferase assay……….…...20
3.6 Gene resequencing panel ……….…………..……21
3.7 Targeted resequencing………..…..….…22
3.8 Targeted resequencing data analysis……….…24
3.9 Statistical analysis………...25
4. RESULTS……….……26
4.1 Genetic screening in familial cases……….….…26
4.2 Genetic screening in sporadic cases………...…27
4.4 Functional effect of the +1521C>G SNP on miR-583 binding……...32
4.5 Case-control study………..…..34
4.6 Linkage Disequilibrium and haplotype analysis………..….36
4.7 High-throughput DNA sequencing………..…37
5. DISCUSSION……….…....39
Mutational screening in sporadic and familial CHDs and a comparison with previous results………..….39
Sequence variations in microRNA targets as genetic risks factors....41
LD and haplotype association testing………..…...42
Novel and rare variants detection by using a targeted resequencing approach………..…43
6. CONCLUSIONS………..……47
