CRISPR overall
CRISPR was first discovered in this type of organism.
Bacteria
What technique do we use to keep cells outside of their natural environment?
Cell culture
This repair mechanism creates indels.
Non-Homologous End Joining
Is germline editing currently allowed in most countries?
no
Name the three components of a nucleotide.
This sequence of nucleic acids is ~20 nucleotides long and is needed to find the place in the DNA to cut.
guide RNA/sgRNA
What biotechnology do we use to insert the Cas9 target DNA into a plasmid?
Recombinant DNA technology
We must also insert THIS into cells if we plan to use HDR.
Donor Template
A scientist in this country produced the first "designer babies"
China
This group is the variable part of an amino acid.
R group/side chain
How many nucleases does Cas9 have?
2
What might we add to a Cas9 plasmid to strengthen confidence in our experiment and be able to sort the cells?
GFP gene sequence
What is a knockout?
A knockout is when we use CRISPR to inactivate a protein in a cell by changing its gene sequence so it no longer works.
What purpose of editing baby genomes is more widely accepted as okay?
Editing to prevent genetic diseases
Name the 4 bases of DNA, how they match up in a double stranded DNA molecule, and the shape that the double stranded DNA molecule makes.
Adenine, Thymine, Cytosine, Guanine
A-T, G-C
Double helix
When CRISPR cuts in another place in the DNA than our target sequence, it is called:
off-target effect
Name two types of transfection we can use to insert the Cas9 and gRNA into cells.
Lipofection and electroporation
This type of CRISPR uses dCas9 to alter one nucleotide pair in DNA.
Base Editing
What is the difference between somatic cell editing and germline editing?
Somatic cell editing only edits cells that are not passed from one generation to the next. Germline cell editing implements changes to genes in the eggs or sperm and these changes could be passed onto future generations.
This type of protein catalyzes biochemical reactions.
Enzyme
What does CRISPR stand for?
Clustered Regularly Interspaced Short Palindromic Repeats
Name two considerations to take into account when designing a sgRNA.
Is there a particular place in the gene we are trying to edit?
Knockout: better to have the edit earlier in the gene
Is there a PAM sequence directly next to the guide RNA?
Will we be aiming for NHEJ or HDR?
Off target effect
What does HDR stand for, and how does it work?
Homology Directed Repair. It uses homologous regions of a donor template with the DNA at the place of the double-stranded break to replace the double-stranded break with the DNA in between the homologous regions.
What is the main argument against the allowance of designer babies?
Deepening the class divide
Explain what a mutation is and a potential consequence.
A mutation is when there is some nucleotide change in the genome, which can lead to a nonfunctional protein.