QuestionQuestion

Immunology
Reading guidelines for paper discussion
CRISPR Cas9 targeting haematoietic stem cells

• What is the overall question that the scientists are trying to address in this paper?
Background research
• What is CRISPR Cas9? Where does it come from and what is it used for?
• What are some examples of B-haemoglobinopathies and the diseases that they cause?
• What causes them?
Introduction
• What is the HBB gene?
• What ways have scientists tried to edit this gene in the past versus what these researchers are proposing?
Methods
In general, what is the approach used by the researchers, such as
• Which cell types are being used and why?
• Where did they get the cells and how?
• What did they do to the cells?
• How can they tell if it worked?
Results:
For each figure (1-4 only) determine the following:
• What is the purpose of this experiment/figure?
• What method was used? (in general- example: used gel electrophoresis to detect the sizes of DNA)
• What does it show? (What is the overall result?)
Discussion:
• What is the overall conclusion?
• What are the implications of this research (i.e. how can it be used in the future?)
• What should be done next?

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Immunology questions

• What is the overall question that the scientists are trying to address in this paper?
Whether there is a safe, viable method for an ex vivo cure for β haemoglobinopathies, by using CRISPR technology to introduce a wildtype HBB gene into patient hematopoietic stem cells.

Background research
• What is CRISPR Cas9?
It is a gene editing technique that utilizes site specific targeting to introduce or delete a region of interest. The complex includes a Cas9 endonuclease, guide RNA specific to your region of interest and PAM sequence.
• Where does it come from and what is it used for?
It originated in bacteria and acted as an adaptive immune system to protect these organisms.
• What are some examples of B-haemoglobinopathies and the diseases that they cause?
Examples include...

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