Lecture 33: CRISPR and Gene Editing Techniques Flashcards
What is Genome editing?
Cutting the genome by DNA scissors, CRISPR/Cas9
What can genome editing be used for?
Removing changing and adding DNA
What is Genomics?
The study of the entirety of an organism’s genes using bioinformatics to find DNA-sequence variations that affect health, disease or drug response
What are Genome-wide association studies (GWAS) used for?
To associate specific genetic variations with particular diseases
What is functional genomics?
A field of molecular biology that attempts to describe a gene (and protein) functions and interactions
What are the differences between the gene editing tools?
They all perform the same function but newer ones are easier to program
What is the advantage to CRISPR-Cas9?
It is easy to design and generate gRNA that binds to target sequence
What were TALENs used to do?
Clone new RVDs
Where was CRISPR/Cas9 found?
In prokaryotes
What does CRISPR/Cas9 do in prokaryotes?
It is part of the prokaryotic immune system that defends against viruses. CRISPR/Cas9 will recognize embedded DNA from viruses and incorporate it in the bacteria’s genome to recognize and inactivate it by cutting it
What is Cas9 recruited to the DNA target site by?
The duplex tracerRNA:crRNA (guide RNA)
What are the three key elements of CRISPR/Cas9?
- the Cas9 DNA endonuclease
- cRNA which contains a 20 bp sequence complementary to the target
- a trans-activating crRNA (tracer RNA) which acts as a bridge between the crRNA and the Cas9 enzyme
What is the cRNA in Crispr?
A 20 BP sequence complementary to the target
What does the trans-activating crRNA (tracrRNA) do?
Acts as a bridge between the crRNA and the Cas9 enzyme
How is DNA endonuclease targeted to a DNA sequence?
Via a single guide RNA (sqRNA) sequence upstream of the PAM
What does DNA endonuclease result in?
A double-stranded break