Genetic Analysis: An Integrated Approach (2nd Edition)
2nd Edition
ISBN: 9780321948908
Author: Mark F. Sanders, John L. Bowman
Publisher: PEARSON
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Chapter 17, Problem 11P
Injection of double-stranded RNA can lead to gene silencing by degradation of RNA molecules complementary to either strand of the dsRNA. Could RNAi (see Sections
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In type II CRISPR-Cas9 systems, what intermediate interacts with the repeat sequences that are added to the guide RNA?
Explain the CRISPR-Cas Mechanism for RNA-Guided Destruction of Invading DNA ?
Prior 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…
Chapter 17 Solutions
Genetic Analysis: An Integrated Approach (2nd Edition)
Ch. 17 - 15.1 What purpose do the bla and lacZ genes serve...Ch. 17 - The human genome is 3109 bp in length. How many...Ch. 17 - 15.3 Ligase catalyzes a reaction between the...Ch. 17 - You have constructed four different libraries: a...Ch. 17 - Using the genomic libraries in Problem 4, you wish...Ch. 17 - The human genome is 3109bp. You wish to design a...Ch. 17 - 15.7 Using animal models of human diseases can...Ch. 17 - 15.8 Compare methods for constructing homologous...Ch. 17 - 15.9 Chimeric genefusion products can be used for...Ch. 17 - 15.10 Why are diseases of the blood simpler...
Ch. 17 - Injection of double-stranded RNA can lead to gene...Ch. 17 - Compare and contrast methods for making transgenic...Ch. 17 - 15.13 It is often desirable to insert cDNAs into a...Ch. 17 - 15.14 A major advance in the s was the development...Ch. 17 - 15.15 The bacteriophage lambda genome can exist in...Ch. 17 - 15.16 The restriction enzymes Xho and Sal cut...Ch. 17 - 15.17 The bacteriophage has a single-stranded DNA...Ch. 17 - 15.20 You have identified a cDNA clone that...Ch. 17 - You have isolated a genomic clone with an EcoR I...Ch. 17 - 15.18 To further analyze the CRABS CLAW gene (see...Ch. 17 - 15.21 You have isolated another cDNA clone of the...Ch. 17 - 15.22 You have identified five genes in S....Ch. 17 - You have generated three transgenic lines of maize...Ch. 17 - 15.24 Bacterial Pseudomonas species often possess...Ch. 17 - 15.25 Two complaints about some transgenic plants...Ch. 17 - 15.26 In Drosophila, lossoffunction Ultrabithorax...Ch. 17 - Prob. 27PCh. 17 - The highlighted sequence shown below is the one...Ch. 17 - The RAS gene encodes a signaling protein that...Ch. 17 - Vitamin E is the name for a set of chemically...Ch. 17 - 15.31 You have cloned a gene for an enzyme that...
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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
- Now 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_forwardDoes one have a practical idea of the role of CRISPR/Cas9 in genome editing via cutting of DNA?arrow_forwardWhat 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_forward
- Propose 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_forwardHow can the bacteriophage T7 promoter be used to controlexpression of a eukaryotic gene in Escherichia coli?arrow_forward. In selecting target cells to receive a transferred gene in gene therapy in an adult person, what factors do you think would have to be taken into account for a successful treatment?arrow_forward
- Which of the following is a correct statement about CRISPR-Cas-9 gene editing? Group of answer choices A single guide RNA (sgRNA) recognizes a genomic region followed by 5'-NGG-3' PAM sequence A single guide RNA (sgRNA) recognizes a genomic region followed by a long sequence palindrome repeat A single guide DNA (sgDNA) recognizes a genomic region followed by 5'-NGG-3' PAM sequence PAM sequences induce single stranded breaks that are then repaired by the CAS-9 enzymearrow_forwardWhat type of RNAi silencing targets DNA? What mechanism does this type of RNAi use? Group of answer choices siRNA; results in methylation of DNA miRNA; results in methylation of the DNA siRNA; results in degradation of DNA asRNA; results in degradation of the DNA asRNA; results in methylation of the DNA miRNA; results in degradation of the DNAarrow_forwardDescribe structural and functional differences between retroviral and non-retroviral transposonsarrow_forward
- Outline the two ways that piRNAs and PIWI proteins inhibit transposable elements.arrow_forwardWhy do researchers believe lncRNA is a promising field for developing drug therapies and CRISPR/Cas can be used to treat disease?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_forward
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