EP INQUIRY INTO LIFE-CONNECT ACCESS
16th Edition
ISBN: 9781260482546
Author: Mader
Publisher: MCG COURSE
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Chapter 26.1, Problem 1AQTC
Summary Introduction
To analyze:
The consequence of unintentional breaks in DNA sequences other than the target sequence due to CRISPR.
Introduction:
CRISPR or Clustered Regularly Interspaced Short Palindromic Repeats are DNA sequences present in prokaryotes. They along with the enzyme Cas9 are part of the defense system of prokaryotes. Now, the CRISPR-Cas9 system is used as a gene-editing tool. It can break DNA strands at the desired location where the gene sequence can be modified according to our requirement.
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With reference to the image below, discuss the process and principle involved for screening/selection of hosts (last stage of cloning) containing the intended recombinant plasmid.
LacZ' = Gene for alpha-peptide of β-galactosidase.
A scientist wants to make a random mutation in a specific gene to investigate that
gene's function. Which of the following methods is she likely to have used to make
this change?
mutagenesis with radiation
random mutagenesis
CRISPR-Cas9 genome editing followed by nonhomologous end joining (NHEJ)
CRISPR-Cas9 genome editing followed by homologous recombination (HR) with a donor
template
oligonucleotide-directed mutagenesis
Gene 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?
Chapter 26 Solutions
EP INQUIRY INTO LIFE-CONNECT ACCESS
Ch. 26.1 - Describe the steps in forming Recombinant DNA.Ch. 26.1 - Discuss how the polymerase chain reaction works.Ch. 26.1 - Prob. 3LOCh. 26.1 - Prob. 1QTCCh. 26.1 - Prob. 2QTCCh. 26.1 - Prob. 1CYPCh. 26.1 - Explain how the PCR reaction amplifies a segment...Ch. 26.1 - Prob. 3CYPCh. 26.1 - Prob. 1AQTCCh. 26.1 - Prob. 2AQTC
Ch. 26.2 - Prob. 1LOCh. 26.2 - Prob. 2LOCh. 26.2 - Prob. 1QTCCh. 26.2 - Prob. 2QTCCh. 26.2 - Prob. 3QTCCh. 26.2 - Prob. 1CYPCh. 26.2 - Prob. 2CYPCh. 26.3 - Compare and contrast in vivo and ex vivo gene...Ch. 26.3 - Prob. 2LOCh. 26.3 - Prob. 1CYPCh. 26.3 - Prob. 2CYPCh. 26.3 - Prob. 3CYPCh. 26.4 - Prob. 1LOCh. 26.4 - Prob. 2LOCh. 26.4 - Prob. 3LOCh. 26.4 - Prob. 1CYPCh. 26.4 - Explain how comparative genomics can provide...Ch. 26.4 - Prob. 3CYPCh. 26 - Prob. S25.1BYBCh. 26 - Prob. S25.2BYBCh. 26 - Prob. S25.3BYBCh. 26 - Prob. 1ACh. 26 - Prob. 2ACh. 26 - The polymerase chain reaction Use RNA polymerase...Ch. 26 - Prob. 4ACh. 26 - Prob. 5ACh. 26 - In this process, cells are removed from the body...Ch. 26 - When a cloned gene is used to modify a human...Ch. 26 - Prob. 8ACh. 26 - Prob. 9ACh. 26 - Prob. 10ACh. 26 - Prob. 1TCCh. 26 - In a genomic comparison between humans and yeast,...
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- What are the major components of the CRISPR-Cas9 system? What mechanism does it employ to combine DNA? Explain the process of how the CRISP-Cas9 system is able to create recombinant DNA. Relate the idea of gene modification to the fields of vaccines and applied microbiology as well.arrow_forwardIt has been shown that infectious agents such as viruses often exert a dramatic effect on their host cell’s genome architecture. In many cases, viruses induce methylation of host DNA sequences in order to enhance their infectivity. What specific host gene functions would you consider as strong candidates for such methylation by infecting viruses?arrow_forwardRestriction sites are palindromic; that is, they read the same in the5' to 3' direction on each strand of DNA. What is the advantage ofhaving restriction sites organized this way?arrow_forward
- Bacteria exposed to viruses incorporate sections of the virus’s DNA into the CRISPR array sequences in their genome. This mechanism allows bacteria to fight off the viruses, like an immune response: the information in CRISPR spacers served as “coordinates” for recognizing and cutting up invading DNA sequences. Describe what might happen under the conditions described after a bacteriophage infects a bacterial cell and releases its DNA into the bacterial cell. Explain why: The cas genes on the bacterial genome contains a frameshift deletion mutation that alters the function of the protein The bacteria will be unable to elicit an immune response and will succumb to the phase infectionarrow_forwardWhat can serve as footprints to identify genome sites that are or have been occupied by transposable elements? options: SNPs Microsatellite DNA Transposase Transposable DNA elements at the insertion sitearrow_forwardDescribe the process of cloning a DNA fragment into theBamHI and PstI sites of the vector pUC18. How would youscreen for clones that contain an insert? and explain the process(steps) by drawingarrow_forward
- Starting with a sample of RNA that contains the mRNA for theβ-globin gene, explain how you could create many copies of theβ-globin cDNA using reverse transcriptase PCR.arrow_forwardA molecular geneticist hopes to find a gene in human liver cells that codes for an important blood-clotting protein. He knows that the nucleotide sequence of a small part of the gene is CTCGACTCACA. Briefly explain how to obtain the desired gene. Briefly describe how to clone the desired gene into a cloning plasmid.arrow_forwardIn order to target a specific region of genomic DNA with CRISPR, researchers must include a guide RNA containing a 20-basepair long spacer sequence that matches the DNA sequence at the target site. (i) How many possible guide RNA spacer sequences are there? (ii) One of the possible risks of genetic engineering methods is “off-target” editing, where a modification of the genome occurs in a part of the genome other than the target site. Imagine you design a 20-basepair guide RNA spacer sequence to target a specific portion of the Zebrafish genome, which is 1.7 billion nucleotides long. Assuming all nucleotides are equally common, estimate the probability that your spacer sequence occurs in at least one other position in the Zebrafish genome.arrow_forward
- Your company has decided to develop a PCR test that will be used to determine Huntington's disease. This genetic test will consist of a single PCR-amplification reaction of genomic DNA that will attach to the disease's gene either side of the repeated region to ensure amplification. Which of the following properties do you need consider when designing the orientation of the two primers relative to the template DNA? a) The primers’ 5′ → 3′ orientations must point toward each other when bound to the template. b) The primers’ 5′ → 3′ orientations must point away from each other when bound to the template. c) The primers’ 5′ → 3′ orientations must go in the same direction when bound to the template. d) The primers must be complementary to each other and to the template.arrow_forwardIn cloning human DNA, why is it necessary to insert the DNA into a vector such as a bacterial plasmid?arrow_forwardThe CRISPR-Cas9 system may be able to be used for somatic gene therapy to treat terminal genetic diseases such as Duchenne Muscular Dystrophy (DMD), which affects 1 in 3,500 male births worldwide. As we talked about in class, DMD results from a nonsense mutation that causes a premature stop codon in Exon 23 of the largest gene yet discovered in the human genome, the 79- exon dystrophin gene. Of the choices below, which would be most likely to restore the protein and muscle function? O A deletion of exon 23 O A deletion of intron 23 O A deletion of intron 23 and exon 23 O An insertion of a corrected exon 23 upstream of the mutant exon 23 O A deletion of exons 23, intron 23, and exon 24arrow_forward
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