gene editing

a. How does the enzyme used in CRISPR-Cas9 differ from enzymes used in recombinant DNA technology?

b.Can you predict a potential risk of somatic cell gene editing?

How to draw a regulatory graph of enzyme glycogen phosphorylase with high ATP and low glucose, with axes labeled, 3 plots on the graph and labeled with given regulators and no regulator present, and plots the correct shape

The following reaction rate data are collected upon analysis of an enzyme catalyzed hydrolysis. The reaction is carried out with two different inhibitors. Determine the modes of inhibition for each inhibitor. (The rates provided are relative to the rate measured in the absence of inhibitor. The enzyme concentration for each set of data is equal.)
No inhibitor
[S]0 (10−2 M) 1.25 3.84 5.81 7.13
v0 0.398 0.669 0.859 1.000
[I1]– –2mmol/L
[S]0 (10−2 M) 1.25 2.5 4.00 5.50
v0 0.179 0.313 0.435 0.529
[I2]– –50mmol/L
[S]0 (10−2 M) 1.75 2.50 5.00 10.00
v0 0.202 0.218 0.240 0.253

Experiment 2- Cloning a DNA Fragment to a Bacterially-Derived Plasmid Vector

“Table 1: Fragment Lengths”

1. What is the expected size of the plasmid plus the cut foreign DNA?

2. What type of ends to the enzymes BamHI and EcoRI produce? How does this type of end facilitate cloning?

3. What enzyme is necessary to permanently link the digested foreign and plasmid DNA together to form the recombinant DNA molecule? How does this enzyme work?

4. How would you clone a gene into a plasmid if there were no common restriction sites between the two DNA sequences?

Studies (kinetic, stereochemical, chemical modification, and site-specific mutagenesis experiments) on the enzyme triosephosphate isomerase have identified the B as Glu165 and HA as His95 (J. R. Knowles, Nature 350, 121 (1991). To aid your understanding, draw the active site as a pocket that extends into the enzyme and place the Flu and His on opposite sides of the pocket. Answer the following questions:

a. Drawing of the pocket:

b. How might the replacement of Glu with Asp affect the catalytic rate?

c. Discuss what form His would be in at a physiological pH of 7.2. Would that pH allow it to act as a general acid? Describe the conditions that allow it to act as a general acid. (hint: what reaction does a Ka refer to?)

Bacterial transformation allows a gene from another organism to be put into a bacterial cell. This would allow the bacterial cell to produce the protein that the gene codes for. Several steps are needed to transform bacteria; put the following steps in the correct order.

1. Add a bacterial promoter and terminator, and add sequences that will be cut by a specific restriction enzyme to the ends of the DNA piece.

2.Cut the cDNA and plasmid with the same restriction enzyme to create sticky ends.

3. Extract mRNA from the eukaryotic cell that expresses the gene.

4. Insert the plasmid carrying the gene from the other organism into the bacterial cell.

5. Produce double-stranded cDNA for the gene of interest using reverse transcriptase.

Medium-Chain Acyl-Coenzyme A Dehydrogenase Deficiency (MCADD)

You’re working in a pediatrics office and a fax comes in that says Baby Smith, a 4-day-old male, has a newborn screen indicating increased risk for medium-chain acyl-coenzyme A dehydrogenase deficiency (MCADD

After familiarizing yourself with MCADD, concisely answer the following questions:

1. What gene is mutated in MCADD and where in the genome is the gene located? (2 pts)
2. Identify the enzyme altered by this mutation and discuss the consequences that result from the impairment of this enzyme. (4 pts)
3. What is the pattern of inheritance for MCADD? (2 pts)
4. How urgent is it to contact this family? Explain (3 pts)
5. Describe the therapeutic approach to this disease. (4 pts)

Imagine that the donut is a sexually reproducing animal and that a single enzyme forms the hole. All donuts observed in the past have had holes. Suppose a mutation occurs in one gene of the donut such that it now codes for an inactive protein instead of the hole-forming enzyme. Which of the following predictions would be most likely in descendants of this mutant donut? For each prediction, explain whether or not it would be a likely prediction.

a. All members of the next generation will have holes

b. Heterozygous donuts will not have holes

c. If the cell in which the mutation occurs undergoes mitosis, subsequent generations of donuts will all have holes

d. The next generation will all be holeless

e.Past generations of donuts have all had holes