Category: Biology

Sickle Cell Anemia: How can we alter the genome to

Question: Present a question. What is the question that you want to answer? (for
example: is it possible to introduce wt CFTR sequence into the mutant CFTR allele and
have it function as CFTR is predicted to?).

2. Introduction: Describe background of gene/condition, etc. What does your audience
need to know about this? What causes it? What is the specific sequence change (if
disease).

How many people are affected? What are the effects? Why is this an
important question to address? [If you are proposing a novel creation, such as GFPkitties, instead describe why this would be a useful or profitable venture, and
how it is similar and different from previous items on the market, like “GloFish”].
be sure to cite sources to describe the background information, including info about the
genes, the role of the genes to be targeted, overview of disease or condition resulting
from mutant alleles (if applicable), or an overview of the function of the genes to be
altered/fused (if applicable).

3. Proposed Strategy: How will you address this? be specific. What will the
constructs look like? describe your strategy for creating the sgRNA construct and
the donor DNA construct, and provide a diagram that includes the relevant
sequence. In your diagram, show: target sequence, PAM, sgRNA sequence,
where mutation occurs (if applicable), where cut site/s is/are. If creating a fusion
protein,  show in your diagram where your inserted gene will go. Also,
show what the sequence of your donor DNA is (no need to include entire gene sequence
for GFP or other inserted protein for fusion protein projects, just indicate the gene with
a labeled rectangle (as in the lab protocols), with the relevant homologous sequence.

4. Delivery: comment on how you would deliver the components needed for
CRISPR to happen, and to which cell population within the organism. Our Moodle site
has a review paper that describes different ways of delivering CRISPR components that
you may find helpful.

5. Results:describe how you would determine if CRISPR was successful.
Describe what the phenotypic effects of the CRISPR would be on the patient or

organism, and also, describe how you would confirm with PCR that the gene edit was
successful. provide the PCR primer sequences.

6. Conclusion: comment on what the overall effects of this new technology would
be on health/society. Put this into context to make it clear in what ways this would be
helpful/useful. In a separate paragraph, discuss drawbacks to this technique, and
importantly, ethical implications for the use of this technology.

What is controversial about CRISPR? Does the technique you are using create heritable changes in DNA?

What about the change you are trying to effect in general – what potential effects does this
have on the community of individuals for whom this would affect (It will be important to

set this up in your introduction section as well –  research how the gene you are
trying edit affects communities with that allele)?

Include figures and legends. The uploaded resources are just guides.