Defective de novo methylation of viral and cellular DNA sequences in ICF syndrome cells
Overview
DNA methylation
CpG distribution
Enzymes
Role of DNA methylation
ICF syndrome
ICF syndrome phenotype
What is known up to now?
Objectives:
Approach
Advantages of EBV-based system
Methods
Bisulfite genomic sequencing
Methylation specific PCR
EBV genome
MSP analysis of Cp and Wp
Expression from Cp and Wp
BGS analysis of Rp
Semiquantative RT-PCR of Zp and Rp
RT-PCR analysis of BHRF1 and BLLF1
Summary of analysis of EBV regions
Scheme of the MAGE-A1 and LAGE-1/2 cellular gene promoters
BGS analysis of MAGE-A1 and LAGE-1/2
Quantification of BGS data
Expression from MAGE-A1 and LAGE-1/2
Summary of the analysis of C-T cellular genes
Schematic of SCP-1 gene promoter
MSP methylation analysis of ICF and normal LCLs for SCP-1
SCP-1 expression monitored by RT-PCR
Expression of DNA methyltransferases in ICF cells
Conclusions
Questions
PPT Slide
References:
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