SECOND SLIDE

 Fragile X Syndrome is a rare genetic disease characterized by intellectual disabilities and

cognitive impairment in mostly males.
 Some distinctive physical features which characterize Fragile X syndrome are a large
head or a long face, a prominent chin, and protruding ears
 The onset of the disease is variable as some cases do not become apparent until puberty
 Motor issues and language delays are also present but become more apparent over
time
 Fragile X syndrome has been found in all major ethnic groups
 Its is more prevalent in men because it is an x-linked dominant disorder. Since females
have two x chromosomes, an inherited damaged FMR1 gene can be compensated by a
healthy FMR1 gene on the other x chromosome. Since males have only one x
chromosome, their inherited damaged copy is their only copy leading it to being more
common in males than females. In fact studies show that 1 in 4000 males are prone to
getting this disease while females have 1-6000-8000 chance in getting this disease
 Currently there are no definitive cures for Fragile X but there are certain treatments
dependent on the symptoms. Behavioral therapy is an available option that helps
manage a range of emotional challenges such as anxiety and anger
 Oxytocin is also administered to individuals, males in particular, who often exhibit
extreme eye gaze avoidance and hyperarousal when they encounter stressful social
situations. Oxytocin is known to alleviate symptoms of social anxiety by lowering cortisol
levels and reducing blood pressure

THIRD SLIDE
 Fragile X Syndrome is caused by a trinucleotide repeat expansion mutation in the FMR1
gene located on the X chromosome at band 27.3 on the q arm
 The disorder is actually called fragile x because some individuals with the disorder were
found to have a segment of their X chromosome that appeared to be broken or fragile
where the FMR1 gene is located
 Usually individuals with Fragile X have a full mutation which means they have over 200
CGG trinucleotide repeats in their DNA while normal FMR1 genes only have about 5-44
repeats
 Individuals with Fragile X have an abnormal methylation and while we know that
methylation often inhibits the expression of certain genes, an abnormal methylation
causes the FMR1 gene to not produce enough FMRP which Is a protein responsible for
normal cognitive development. This is a cause of the intellectual disabilieites, cognitive
impairment, etc.
 There are cases of premutations in which the DNA has only 55-200 CGG repeats
 Individuals with the premutations do not have fragile x syndrome but they do run the
risk of having other FMR1 disorders such as Fragile X-associated ataxia syndrome which
is a late onset neurodegenerative didorder, usually after the age of 50
 Premutations may become full mutations when passed from generation to generation
but the risk is dependent on whether the premutation carrier is a male or female
 Females with a premutation are at risk of having children with Fragile X syndrome
because the number of CGG repeats can increase as the gene is passed on from
generation to generation
 Male carriers who have male offspring have no risk to pass on the premutation because
they do not contribute an X chromosome to their sons
 Meanwhile female offspring of male carriers always inherit the premutation and
therefore, the grandchildren of male carriers are at risk of getting fragile X
FOURTH SLIDE
 Two types of genetic testing are used to investigate whether an individual has Fragile X
syndrome or not. The southern blot determines if the gene has had a full mutation. An
FMR1 gene probe which is a single stranded copy of the FMR1 gene is used to read the
isolated DNA fragments. PCR determines the actual number of CGG repeats. It is much
less expensive and quicker than a southern blot but it is not the test of choice to
diagnose a full mutation because it does not test for dna methylation. It does however
determine smaller premutations and is helpful in carrier testing. PCR uses dna
polymerase to make millions of copies of the short section of the patients frm1 gene
containing the repeat. When its transferred onto a gel, pieces differing in length by as
little as one CGG repeat can be distinguished.

FIFTH SLIDE
 Let me give you a very brief summary of the case study. Melissa has a brother named
David who has fragile X syndrome. Melissa is married to Paul and they get pregnant.
Melissa calls her mom, Emma, to tell her the good news but Emma is afraid the baby will
have Fragile X. Melissa and her brother make an appointment to get genetic testing
done on both of them and on the fetus. A southern blot was conducted.

SIXTH SLIDE
 If we look at The first 3 bands, we see that they are homozygous controls for a full
mutation, normal DNA, and a premutation resepctively.
 The fourth band is homozygous and lines up with the full mutation band.
 Bands 5,6,and 7 are heterozygous because they have an allele that’s lines up with
normal DNA and another allele lines up with the premutation control.
 The fragments are of varying length based on the allele.
SEVENTH SLIDE
1. What is the chance that Melissa is a carrier?
 There is a 25% that Melissa is a carrier. Being that the disease is x-linked, the
mother is definitely a carrier as she was able to pass it onto her son.
 That leaves a daughter that maybe a carrier and an unaffected son and an
unaffected daughter.

2. What conclusions can you draw from the results of the DNA analysis?
 Emma, Melissa, and the baby are heterozygous for Fragile X syndrome because
their samples all aligned with the normal DNA control, however they do have
wider bands which represent that they have a premuated allele and a normal
allele, making them carriers.
 David is homozygous for Fragile X because he has a single band that aligns very
closely with the full mutation band.
3. How would you counsel Melissa based on the results of her test?
 I would provide a risk assessment for Melissa saying that her offspring has a 50%
chance of inheriting Fragile X syndrome due to the fact that she is a carrier.
 I would also take a full family history to determine if the baby is at risk for any
other genetic disorders.
 Further into pregnancy like about 10-12 weeks, I would advise a Chorionic villus
sampling which checks cells from the placenta for the FMR1 gene.
 About 15 to 18 weeks into the pregnancy, I would advise a Amniocentesis test
which checks the amniotic fluid for the FMR1 gene.

4. What issues may be raised by the results of the testing?

 Because the blot of the fetus is identical of to the mother and the grandmother,
it can be assumed that the fetus will be a carrier for Fragile X syndrome.