Genetics: From Genes to Genomes
6th Edition
ISBN: 9781259700903
Author: Leland Hartwell Dr., Michael L. Goldberg Professor Dr., Janice Fischer, Leroy Hood Dr.
Publisher: McGraw-Hill Education
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Chapter 18, Problem 24P
| a. | Figures 18.9 and 18.12 demonstrated methods to produce mouse knockouts and knockins, respectively.CRISPR/Cas9 can make the same knockouts and knockins, and most mouse geneticists would now choose to use this new technology instead of the other methods. Explain why, respectively CRISPR/Cas9 is an easier and more efficient way to perform targeted mutagenesis in mice. |
| b. | How could you use the CRISPR/Cas9 technique to obtain a conditional knockout of a gene’s function only in a specific tissue of a multicellular organism?  |
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a. Describe two ways you could potentially make atransgene that would inhibit the function of a specific gene in a transgenic organism. (Hint: For oneof these techniques, recall the discussion of RNAinterference in Chapter 17.)b. Discuss how you could use either of these methodsto construct a mouse model for a recessive humangenetic condition associated with a loss of function, such as cystic fibrosis.
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
Describe three possible uses of site-directed mutagenesis.
Chapter 18 Solutions
Genetics: From Genes to Genomes
Ch. 18 - Match each of the terms in the left column to the...Ch. 18 - Mice are usually gray, but a mouse geneticist has...Ch. 18 - Sometimes, genes transferred into the mouse genome...Ch. 18 - In mice, a group of so-called Hox genes encode...Ch. 18 - The fly eyes shown in Fig. 18.7 are malformed...Ch. 18 - This problem concerns a technique called enhancer...Ch. 18 - Fish and other organisms that live in the Arctic...Ch. 18 - a. Describe two ways you could potentially make a...Ch. 18 - Figure 18.6 shows a picture of Glofish ,...Ch. 18 - Some people are concerned about the possible...
Ch. 18 - The goal of the Knockout Mouse Project is to...Ch. 18 - Prob. 12PCh. 18 - Prob. 13PCh. 18 - a. Which genome manipulation technique would you...Ch. 18 - a. Diagram a knockin construct that could have...Ch. 18 - Prob. 16PCh. 18 - Prob. 17PCh. 18 - The transcription factor Pax6 is required...Ch. 18 - Mouse models for human genetic diseases are...Ch. 18 - One way to determine where inside a cell a protein...Ch. 18 - In Problem 5 in Chapter 17, you saw that a SNP...Ch. 18 - Scientists now routinely use CRISPR/Cas9 to make...Ch. 18 - Geneticists are currently considering using...Ch. 18 - a. Figures 18.9 and 18.12 demonstrated methods to...Ch. 18 - Nonhomologous end-joining NHEJ of a double-strand...Ch. 18 - One problem that researchers sometimes encounter...Ch. 18 - Researchers at the University of California at San...Ch. 18 - Prob. 28PCh. 18 - F. Port and S. Bullock at the University of...Ch. 18 - On Fig 18.14, locate the PAM site and identify the...Ch. 18 - Prob. 31PCh. 18 - Prob. 32PCh. 18 - Recall that Leber congenital amaurosis LCA, a form...Ch. 18 - One potential strategy for gene therapy to correct...Ch. 18 - Recently, scientists have used a mouse model for...
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Need a deep-dive on the concept behind this application? Look no further. Learn more about this topic, biology and related others by exploring similar questions and additional content below.Similar questions
- Explain 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_forwardAn EMS mutagenesis screen identified several mutants in this gene. The figure shows the position of two of these mutations a and b. The ucleotides are altered in these 2 different swo-1 mutant alleles. Use the genetic table to describe any AA changes. Name the type of mutation and describe its effect on swo-1 mRNA and protein for each of the mutations 5. The swo-1 a mutation leads to worms with more body wall muscle, whereas worms with the swo-1 b mutation are not able to move. Based on these phenotypes and the findings from questions 3 and 4, describe the role thewild-type version of this protein plays in muscle function.arrow_forwardYou would like to study the effect of GeneX in the developing mouse liver. However, your supervisor says that GeneX knockouts (loss of function mutants) are embryonic lethal. What technique could you use to knockout GeneX only in the liver without causing embryonic lethality. Describe how you would do this experimentarrow_forward
- siRNA and CRISPR/Cas9 each interfere with gene expression by a sequence-directed mechanism. A. Identify the target macromolecule (DNA, RNA or Protein) of a specific siRNA/RISC andCRISPR/Cas9 complexsiRNA:CRISPR/Cas9: B. If a scientist would like to permanently turn off expression of a particular human gene inthe liver for the lifetime of an individual, which of these techniques would bepreferable? Briefly explain whyarrow_forwardNow that you understand how the CRISPR-Cas9 system works, think back to the experiments discussed in the introduction to this chapter, in which researchers used CRISPR-Cas9 genome editing to treat mice with Duchenne muscular dystrophy. Why did the researchers choose to cut out the entire exon 23 in the mice with the disorder? Why not replace the specific mutation using a donor piece of DNA and homologous recombination? Propose some possible explanations.arrow_forwardGene expression can be disrupted by techniques such as homologous recombination and RNA interference. What is the functional difference between these two methods in terms of the ultimate effect on gene expression? Why might homologous recombination or an alternative genome-editing method (e.g., CRISPR/Cas9) be preferred over RNA interference?arrow_forward
- Researchers have identified a gene (FR) responsible for watermelon resistance to infection by Dacus curcurbitae (a close relative of Drosophila melanogaster). They isolate RNA from resistant (FR+) and sensitive (fr-) watermelons and use a probe that will recognize both FR+ and fr- transcripts. They also isolate protein from resistant and sensitive watermelons and perform a Western blot using an antibody that can recognize the fr- and FR+ protein. Describe the results illustrated below and give a plausible molecular explanation for these observations.arrow_forwardPropose some specific uses of a modified CRISPR-Cas system as a general RNA-guided device for altering cellular functions. What might these functions be, and how could CRISPR-Cas be used to study them?arrow_forwardThe restriction digests of the cloned Drosophila gene can provide direct visible evidence of a mutation, as these samples come from a clone of the gene. In order to similarly detect a mutation in the copy of the endogenous gene within the Drosophila genome, a mechanism for specifically detecting restriction fragments from that gene among the complex set of fragments generated in a restriction digest of the entire Drosophila genome. Remember that the Drosophila melanogaster genome consists of ~123,000 kb. For a 10.2 kb Drosophila gene, what fraction of the genome does this gene constitute?arrow_forward
- List three possible uses of site-directed mutagenesisarrow_forward. a. Mice engineered to block expression of the geneencoding the telomerase protein age at a muchfaster rate than normal and have decreased lifespans. When expression of the telomerase proteinis turned back on in mice that are prematurely old,many negative effects of this aging are rapidly anddramatically reversed. Provide a possible explanation for these results.b. The results of these mouse experiments have ledsome researchers to propose that treatments thatcould lead to overexpression of the telomerase genemight serve as a “fountain of youth” leading to reversal of aging in humans. Why do you think weshould be very cautious about trying such treatments? Your argument should address why it mightbe advantageous to multicellular organisms for mostof their somatic cells not to express telomerase.arrow_forwardA series of exonuclease deletions were used to study the promoter of the rice hemA gene, giving the results shown below (the intact promoter is on top). Based on these results, what conclusions could you draw from each of the deletion construct, and what do you know about the nature of the promoter? Relative activity -500 Reporter gene 100% Reporter gene 180% -350 -250- Reporter gene 180% 75% -150 Reporter gene 45% Reporter gene -100 0% - 8 Reporter genearrow_forward
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