6-BSC Hypertrophic Cardiomyopathy and Linkeropathies

Question 1

What are linkeropathies?

Answer 1

inablilty or loss of enzyme that links sugars to proteoglycans (found in ECM)

Question 2

Whole exome sequencing in the detection of new genetic variants?

Answer 2

have to look to many sources and match up everything to the symptoms.

once you find the mutation, what does it do?

look at mRNA, protein and activity to see if it matches up.

Question 3

Describe pathophysiology of variants in the gene B3GAT3

is there normal mRNA, protein, and activity levels?

Answer 3

B3GAT3 (gene) –> GLCATI (enzyme)

patient makes same amount of mRNA but less protein than control. 3% of activity of WT

linkeropathies should be considered in dd with hypotonia, joint laxity, congenital heart disease and fractures

Question 4

What gene should we focus on for hypertrophic cardiomyopathy?

Answer 4

MYBPC7

Question 5

What gene should we focus on for hypertrophic cardiomyopathy?

How is it related to sarcomere unit?

Answer 5

MYBPC7

Dysfunction in contraction related to reuptake of calcium

Question 6

What gene should we focus on for hypertrophic cardiomyopathy?

How is it related to sarcomere unit?

Answer 6

MYBPC3

Dysfunction in contraction related to reuptake of calcium

Question 7

What is Cas-9 job?

And what other players are critical for targeting specific DNA sequence?

Answer 7

Cas-9= nuclease, elicits ds breaks
sgRNA= single guided, takes Cas-9 to bring nuclease to specific site on DNA  

Cas-9 will recognize specific sequences upstream of PAM
–PAM- specific 3 nucleotides.

Question 8

What doe the ssDON do?

Answer 8

single stranded deoxyoligonucleotide

Serves as template with homologous recombination

Question 9

What stage in the cell cycle were the oocytes in when they were injected with CRSPR Cas9?

Answer 9

S phase

fertilized with heterozygous sperm, equal Mutant and WT

Question 10

What findings did they find about which systems were used?

Answer 10

Embryos preferentially utilized HDR
And used WT allele as template, no sgDON

Embryos choose more accurate DNA repair

Efficiency was way higher in embryos then patient induced pluripotent stem cells.

Question 11

How did they increase the efficiency? make less mosaicism in embryos?

Answer 11

Changed to injecting a cell in M phase instead of S phase. Chang the alleles before they replicate.

Question 12

What is PGD (related to designer babies)

Answer 12

PGD- preimplantation genetic diagnosis