Genetics: Analysis and Principles
6th Edition
ISBN: 9781259616020
Author: Robert J. Brooker Professor Dr.
Publisher: McGraw-Hill Education
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Textbook Question
Chapter 17, Problem 4EQ
As described in Chapter 21, the CRISPR-Cas system has been modified so it can be used as a gene mutagenesis tool (look ahead to Figure 21.13). Describe how the gene mutagenesis tool works, and explain how the natural CRISPR-Cas system is altered to produce this tool.
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List three possible uses of site-directed mutagenesis
i) Suppose we want to insert the GFP sequence after the promoter of a gene X to create a fusion protein. Please describe the strategy that you will follow to edit this gene with CRISPR-Cas9 technology, in order to visualize the expression of gene X in live cells.
(ii) Please explain how a knockout mouse for a gene X can be produced with CRISPR-Cas9 technology
There are very different rules governing genome edited crops in the European Union and in the United States of America, according to Cohen. Explain why CRISPR-edited crops could be considered to be a type of GMO (per the EU) and why they could be considered equivalent to naturally occurring mutations (per the USA). In your explanation be sure to (1) discuss how CRISPR constructs are currently introduced into plants and (2) how all evidence of the CRISPR apparatus/process can be eliminated from the genome, leaving only the edited sequence(s).
Chapter 17 Solutions
Genetics: Analysis and Principles
Ch. 17.1 - Which of the following can bind to ncRNAs? a. DNA...Ch. 17.1 - 2. When an ncRNA functions as a decoy, it
a....Ch. 17.1 - Prob. 3COMQCh. 17.2 - 1. Which of the following functions does HOTAIR...Ch. 17.3 - 1. The process of RNA interference may lead to
a....Ch. 17.3 - 2. In catalyzing the methylation or...Ch. 17.4 - 1. Which of the following is a function of SRP?...Ch. 17.5 - 1. Which of the following components are needed...Ch. 17.5 - 2. In the CRISPR-Cas system, what does the...Ch. 17.5 - Which of the following is a function of the...
Ch. 17.6 - Prob. 1COMQCh. 17.6 - Let's suppose that the overexpression of a...Ch. 17 - 1. List and briefly describe four types of...Ch. 17 - 2. An ncRNA may have the following functions:...Ch. 17 - 3. What is meant by the term RNA world? Describe...Ch. 17 - Explain how HOTAIR plays a role in the...Ch. 17 - What is the phenomenon of RNA interference (RNAi)?...Ch. 17 - With regard to RNAi, what are three possible...Ch. 17 - 7. What is the difference between an miRNA and an...Ch. 17 - Together with a specific set of proteins, snoRNAs...Ch. 17 - Describe the structure of SRP in eukaryotes, and...Ch. 17 - Look at Figure 17.6 and predict what would happen...Ch. 17 - Compare and contrast the roles of crRNA and...Ch. 17 - In the CRISPR-Cas system, does the tracrRNA act as...Ch. 17 - Prob. 13CONQCh. 17 - Outline the steps that occur when piRISCs silence...Ch. 17 - List five types of cancer in which ncRNAs can be...Ch. 17 - Prob. 16CONQCh. 17 - A protein called trypsin, which plays a role in...Ch. 17 - Prob. 2EQCh. 17 - Prob. 3EQCh. 17 - As described in Chapter 21, the CRISPR-Cas system...Ch. 17 - Prob. 5EQCh. 17 - Prob. 6EQCh. 17 - Prob. 1QSDCCh. 17 - Go to the PubMed website and do a search using the...
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- in the cloning vector, what would be the plausible impacts if a mutation at the Ori site renders it non- functional?arrow_forwardHow to incorporate prokaryotic gene into eukaryotic cell to make an transgenic organism?In this case what promoter will be used.explain the process in detailsarrow_forwardMutagenesis is a technique in which genetic information of an organism is altered in a stable manner resulting in a mutation. It may occur spontaneously in nature of as a result of exposure to mutagens. It can also be achieved experimentally using optimized laboratory procedures. (i) (ii) What is site directed mutagenesis (SDM)? Explain how SDM can assist in the integration of a His-tag at the end of your gene of interest.arrow_forward
- Suppose a researcher previously cloned gene Y into M13 bacteriophage vector. Gene Y encodes a product called peptide Y. A region of gene Y contains the DNA sequence ATG-CGC-GAA-CTG-GTG-AAC-TAA. The researcher wishes to change a Val residue to an Ala residue in this region of peptide Y using site-directed mutagenesis. What should be the sequence of the mutant oligonucleotide primer in this region? You may use a codon table. mutant oligonucleotide primer sequence: GGC-GGC-GAA-CTG-GTG-AAC-TAA Incorrectarrow_forwardPrior to the publication of the article from which this assay comes from (picture), the Cas9 endonuclease was known to be involved in CRISPR RNA (crRNA) maturation, but the mechanism by which Cas9 participates in targeted DNA disruption was unknown. To test whether and how Cas9 might be able to target DNA cleavage, researchers developed an in vitro assay. One of the first steps was to purify the Cas9 protein from Streptococcus pyogenes and several orthologs. The description of the figure states: SDS-PAGE analysis of purified Cas9 orthologs. Cas9 orthologs were purified as described in Supplementary Materials and Methods. 2.5 ug of each purified Cas9 were analyzed on a 4-20% gradient polyacrylamide gel and stained with SimplyBlue Safe Stain. Answer the following questions based on this information and the figure. a. What is the experimental/research question and state a hypothesis related to this question. b. What is the approach of the figure and provide a description of techniques…arrow_forwardAlthough a large number of mutagenic chemicals are known,none is known that induces mutations in only a single gene(gene-specific mutagenesis). From what you know aboutmutagens, explain why it is unlikely that a gene-specificchemical mutagen will be found. How then is site-specificmutagenesis accomplished?arrow_forward
- You are examining the processing of rRNA in E. Coli using a Northern blot. Normally, a 30S rRNA transcript is transcribed and then processed (cleaved) into 23S, 16S, and 5S mature rRNAs. You suspect the gene RPO7 is involved, so you make a mutant strain of bacteria and compare it to a Wild-type (Wt) strain. You run RNA extracts from the two strains on a Northern blot and probe it with a radioactive probe that binds to all rRNA. You get the following results.arrow_forwardExplain how site directed mutagenesis can be accomplished using M13 bacteriophage. Using this method often results in only 5% of plaques with mutant gene. Explain how the procedure has been modified to enrich for the number of plaques with the mutant gene.arrow_forwardNorthern blots are valuable tools to analyze the mRNA level present in a sample. Describe an experiment that uses the Northern blot as a tool and how you would visualize the mRNA giving at least two examples of probe types.arrow_forward
- Outline the three stages of CRISPR-Cas action.arrow_forwarda gm animal that may be approved for human consumption by the time this book is published is a super muscly pig made by inactivation of the myostatin gene. During normal development, myostatin protein prevents the overgrowth of muscles. (Given that effective deletions can be made in the genome by using CRISPR/Cas9. It targets deletions of varied length. By this technique, the transcriptional status of the target gene is not affected. The gene deletions produced by CRISPR/Cas9 lead to the production of the correct type of junctions at very high frequencies), how could the super muscly pig have been generated? Answer the questions in the photo as well. ( i included my answer for 22a in the question above, if you believe you have a better answer by all means include that as well)arrow_forwardIn the bacteriophage T7 system used to express recombinant proteins, the gene of interest is fused to T7 promoter and T7 RNA polymerase is separately cloned into the same cell. What is the main reason this system uses T7 RNA polymerase instead of relying on the bacterial RNA polymerase? To restrict the expression of bacterial protein expression To enhance the amount of recombinant protein expression To enhance the expression of bacterial protein expression To restrict the amount of recombinant protein expression To enable the expression of T7 viral protein expressionarrow_forward
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