List and briefly describe all essential components of CRISPR-Cas9 system
List and briefly describe all essential components of CRISPR-Cas9 system
Homework Answers
CRISPR ( Clustered regularly interspaced short palindromic repeats) is a family of DNA sequences found in the genomes of prokaryotic organisms like bacteria and archae. The CRISPR-Cas9 system isa prokaryotic immune system that confers resistance to foreign elements such as those present in plasmids and phages.
CRISPR-Cas9 system contains two essential components- a guide RNA (gRNA or sgRNA) and a CRISPR associated endonuclease (Cas9 protein).
(a) Guide RNA (gRNA):- The gRNA is a short synthetic RNA composed of a scaffold sequence necessary for Cas-binding and a user defined 20 nucleotide spacer that defines the genomic target to be modified.
(b) Cas9:- It is a protein that uses CRISPR sequences as a guide to recognise and cleave specific strands of DNA that are complementary to the CRISPR sequence.
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List and briefly describe all essential components of
CRISPR-Cas9 system
3. List similarities and differences between RNA interference and CRISPR-Cas9 system.
3. List similarities and differences between RNA interference and CRISPR-Cas9 system.
8) Although the CRISPR/Cas9 system has been widely used to edit genomes, as discussed in class...
8) Although the CRISPR/Cas9 system has been widely used to edit genomes, as discussed in class this technology has been repurposed for numerous other applications. Briefly describe how this technology is being used to activate gene expression.
4. The CRISPR-Cas9 system is an important new technique in molecular biology. What is the natural...
4. The CRISPR-Cas9 system is an important new technique in
molecular biology. What is the natural function of this system?
Describe how you would use this system to generate a null mutation
in another organism (i.e. explain Figure 6-43). How does it work?
What is the modification of the method that allows for correction
of a mutation (e.g. the mouse crystalline gene)? And lastly, what
are the problems with the CRISPR system?
FIGURE 6-43 Single-nucleotide mutations
can be introduced into...
Copy of In the CRISPR/Cas9 genome editing system, the job of the guide RNA is to......
Copy of In the CRISPR/Cas9 genome editing system, the job of the guide RNA is to... o cut the target sequence The job of the guide RNA is not fully known yet. o identify the correct sequence to be cut o help the plasmid enter the cells hide the plasmid from the immune system
CRISPR-Cas9 provides an unprecedented means of modifying the genome of almost any organism. Which of the...
CRISPR-Cas9 provides an unprecedented means of modifying the genome of almost any organism. Which of the following component(s) of this system provide(s) the specificity of the endonucleolytic cleavage? the sgRNA the PAM sequence Cas9 a and b a, b and c
Briefly describe the seven components of clinical integration and give reasons why each is essential. In...
Briefly describe the seven components of clinical integration and give reasons why each is essential. In your opinion, which one is the hardest component to develop and manage successfully?
Which of the following statements is FALSE? In the context of CRISPR-Cas9 gene editing system, the...
Which of the following statements is FALSE? In the context of CRISPR-Cas9 gene editing system, the PAM sequence: a. A sequence adjacent to the target DNA sequence to be cleaved b. Is present on the non-complementary target DNA strand to the crRNA c. Has the sequence “5’-NGG-3’” where N is any nucleotide d. Stands for "protospacer adjacent motif" e. Is present in the bacterial genome
Question 7 (1 point) CRISPR/Cas9 is often used to make mutations in zebrafish. Which statement is...
Question 7 (1 point) CRISPR/Cas9 is often used to make mutations in zebrafish. Which statement is most correct? Capping the sgRNA is essential for its stability. The Cas9 mRNA is is used to target the native Cas9 protein to the locus to be mutated. the sgRNA uses DICER to cut up a the twenty nucleotide target region of the DNA, starting at the 3-prime end. O neomycin is used to select the cells that inherit the Cas9 insertion. O the...
All of the following are required for genome editing with the CRISPR-Cas 9 system except: A....
All of the following are required for genome editing with the CRISPR-Cas 9 system except: A. Cas9 nuclease B. An sgRNA C. A repair mechanism for a double-strand DNA break (DSB) D. Invading DNA protospacers E. A PAM sequence
You wish to edit a gene in a population of stem cells using CRISPR/Cas9. You design...
You wish to edit a gene in a population of stem cells using CRISPR/Cas9. You design a plasmid and transfect it into the cells. Your plasmid included the guide RNA sequence with promoter, the Cas9 gene inserted between an appropriate promoter and termination sequence, and the usual Origin of Replication and Resistance/Marker gene. However, instead of being edited, your target gene is silenced and its protein is not produced at all. What is the most likely explanation? The cells have...
4. The CRISPR-Cas9 system is an important new technique in
molecular biology. What is the natural function of this system?
Describe how you would use this system to generate a null mutation
in another organism (i.e. explain Figure 6-43). How does it work?
What is the modification of the method that allows for correction
of a mutation (e.g. the mouse crystalline gene)? And lastly, what
are the problems with the CRISPR system?
FIGURE 6-43 Single-nucleotide mutations
can be introduced into...
Copy of In the CRISPR/Cas9 genome editing system, the job of the guide RNA is to... o cut the target sequence The job of the guide RNA is not fully known yet. o identify the correct sequence to be cut o help the plasmid enter the cells hide the plasmid from the immune system
Question 7 (1 point) CRISPR/Cas9 is often used to make mutations in zebrafish. Which statement is most correct? Capping the sgRNA is essential for its stability. The Cas9 mRNA is is used to target the native Cas9 protein to the locus to be mutated. the sgRNA uses DICER to cut up a the twenty nucleotide target region of the DNA, starting at the 3-prime end. O neomycin is used to select the cells that inherit the Cas9 insertion. O the...
You wish to edit a gene in a population of stem cells using CRISPR/Cas9. You design a plasmid and transfect it into the cells. Your plasmid included the guide RNA sequence with promoter, the Cas9 gene inserted between an appropriate promoter and termination sequence, and the usual Origin of Replication and Resistance/Marker gene. However, instead of being edited, your target gene is silenced and its protein is not produced at all. What is the most likely explanation? The cells have...
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