a) Why do we need to alter the sequence of the PAM or the Crispr in...
a) Why do we need to alter the sequence of the PAM or the Crispr in our repair template?
b) Does the tag have to be inserted where the CAS9 cuts the DNA?
Homework Answers
We need to alter sequence of Pam or crisper sequence in our repair template so that it is not targeted for cutting again and again by crisper cas9 system. The change in pam make these sequences unrecognised once after the required editing already took place.
B) Yes! florescent tag ( a foreign dna sequence in a endogenous DNA) protein has to be inserted at the place where DNA editing has been carried.
Add Answer to:
a) Why do we need to alter the sequence of the PAM or the Crispr
in...
4. What is the function of each component of the CRISPR-Cas9 system? (1.5 pt) PAM site:...
4. What is the function of each component of the CRISPR-Cas9 system? (1.5 pt) PAM site: Guide RNA: Cas9: 5a. In the DNA below, the "I" line indicates where there is a mutation in a gene. Design a gRNA to find this mutation. Your gRNA will be 8 bases long and will not include the PAM sequence. Keep the following in mind: [2 pts total] i. Scan the top strand left to right and find the PAM site. ii. When...
please help answer these questions 3. What are the similarities, differences, advantages, and disadvantages of CRISPR-based...
please help answer these questions
3. What are the similarities, differences, advantages, and disadvantages of CRISPR-based gene editing versus Zinc-finger nucleases and TALENS? 4. What is crRNA and what does it do? 5. What is tracrRNA and what does it do? 6. What is the PAM sequence and what is its significance? 7. What can nuclease-deficient Cas9 (acas) proteins be used for? 8. You want to insert DNA encoding an epitope tag to the end of a specific gene you...
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
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
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...
We know that two things must be recognized by Cas9 before it will perform cleavage: (1)...
We know that two things must be recognized by Cas9 before it will perform cleavage: (1) the correct PAM sequence, and (2) adequate base pairing between the spacer sequence of the RNA and the candidate DNA target. We also know that PAM recognition happens first. Does this make sense? In other words, if you were designing an enzyme that had to do these two tasks (in the same way that Cas9 does them), would you choose to test for PAM...
1. What is the source of the “spacer” DNA in the bacterial CRISPR locus? 2. Why...
1. What is the source of the “spacer” DNA in the bacterial CRISPR locus? 2. Why does the cas9 enzyme have two nuclease domains? 3. What is the relationship between the tracrRNA and the regularly interspersed repeats of the CRISPR locus? Why is this important? 4. In the lab version of the CRISPR/cas9 system, what is the “chimeric RNA”?
Please help me answering these multiple questions of how the CRISPR system works, thank you! 1....
Please help me answering these multiple questions of how the CRISPR system works, thank you! 1. Match the CRISPR component with its function. (matching the upper case letters to the lower case letters) A). Cas9 B). gRNA (sometimes called sgRNA) C). Non-homologous End Joining (NHEJ) D). Homology-directed Repair (HDR) a). A short RNA molecule that guides a DNA cutting enzyme to a specific location in genomic DNA. b). A type of DNA repair mediated by a set of enzymes all...
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...
1. what is CRISPR gene editing? 2. what enzyme is used to cut DNA? 3.Why do...
1. what is CRISPR gene editing? 2. what enzyme is used to cut DNA? 3.Why do bacteria use CRISPR? 4.How is the enzyme targeted to a specific DNA sequence? 5.what need to be done in order to make sure the proper human protein is made in bacteria cells? 6.How can you check for proper orientation of the inserted DNA in the plasmid? 7. Why are iPSCs useful for studying hereditary neuronal diseases such as Alzheimer's. 8. How are pig cells...
4. What is the function of each component of the CRISPR-Cas9 system? (1.5 pt) PAM site: Guide RNA: Cas9: 5a. In the DNA below, the "I" line indicates where there is a mutation in a gene. Design a gRNA to find this mutation. Your gRNA will be 8 bases long and will not include the PAM sequence. Keep the following in mind: [2 pts total] i. Scan the top strand left to right and find the PAM site. ii. When...
please help answer these questions
3. What are the similarities, differences, advantages, and disadvantages of CRISPR-based gene editing versus Zinc-finger nucleases and TALENS? 4. What is crRNA and what does it do? 5. What is tracrRNA and what does it do? 6. What is the PAM sequence and what is its significance? 7. What can nuclease-deficient Cas9 (acas) proteins be used for? 8. You want to insert DNA encoding an epitope tag to the end of a specific gene you...
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...
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