Fragile X Syndrome Mutation by Methylation Sensitive PCR
1. Detection of Fragile X Syndrome Mutation using By: Connie Ross MS-PCR
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7. Diagnostic Methods A good test must have the ability to determine (both) Expansion & Methylation
8. XIST Gene as a Control FMR1 XIST FMR1 XIST FMR1 XIST Male Female Inactive Active XIST Functions as an Internal Standard & Control in the Promoter Region to Determine Methylation Its Methylation Pattern Opposes that of FMR-1 X Y X a X i CH 3
14. 100 bp marker Normal Female Normal Female Fragile X Female Neg. Ctrl Normal Male Fragile X Male 100 bp marker Promoter Region MS-PCR 100 bp 200 bp 300 bp 400 bp
15. Re: Methylation Other Normal Samples Tested 100 bp marker Male - H0011 FM Std. - H0016 Male - H0012 Male - H0014 Neg. Ctrl 100 bp marker 100 bp marker Female - H0020 Female - H0021 Female - H0022 Female - H0023 Female - H0024 Female - H0025 FM Std. - H0016
22. Acknowledgements Dr. Nader Ghebranious PhD Lynn Ivacic Chuck Dokken Dr. Philip Giampietro MD Dr. E. McPherson MD Christina Zaleski Steve Kaiser Dr. Mark Borchardt PhD Research Foundation Donors
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Editor's Notes
“ The objective of my project was to develop an efficient Dx test for Fragile X syndrome based on a later sited publication”
Most common cause of inherited mental retardation in males “ most common phenotypic characteristic…”
“ X-Inactive Specific Transcript” (XIST) XIST does not produce a protein - Causes chromosome inactivation Expression of XIST causes…
Low (but detectable) FMR-1 gene expression
By Formula: Would have to be less than 229 bp @ RU & less than 285 bp @ RM to have less 55 repeats capable of producing a PCR product Avg ~29-CGG = 151bp
In a clinical setting, will have to be performed with fluorescent markers using an ABI sequencer
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