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?

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?

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter16: Reproductive Technology, Genetic Testing, And Gene Therapy

Section: Chapter Questions

Problem 12QP: In selecting target cells to receive a transferred gene in gene therapy, what factors do you think...

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Which two molecules are necessary for every CRISPR gene editing experiment? What additional molecule is needed for gene editing using homology directed repair (HDR) that is not needed for non-homologous end joining (NHEJ)? What methods are used to introduce these molecules into the cells that are being targeted for gene editing?
What is homologous recombination and where/when does it typically take place in an organism. Homologous recombination is also used experimentally during gene targeting. If you wanted to remove the function of the mouse gene fibroblast growth factor 4 (FGF4) using gene targeting by homologous recombination, what are the different aspects of a targeting vector you would need to make?
In 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 expression
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.
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.
What is the difference between nonhomologous end-joining (NHEJ) and homology-directed repair (HDR) in the context of genome editing?
Codon optimization is a widely used process for recombinant expression in prokaryotic systems. Regarding this process, mark the incorrect alternative. * A)increases the overall yield of expression of recombinant proteins. B)prevents the formation of truncated proteins. C)increases the frequency of tRNAs that recognize rare codons. D)does not interfere with the primary sequence of the recombinant protein.
In selecting target cells to receive a transferred gene in gene therapy, what factors do you think would have to be taken into account?
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
CRISPR-Cas9 is used by bacteria as a type of immune system against invading viruses. In recent years, it has been developed into a revolutionary tool that can induce double-stranded breaks at very specific locations in DNA in eukaryotic cells. CRISPR-Cas9 activity typically results in indel mutations in the targeted gene. Which DNA repair system most likely caused these indels? a) non-homologous end-joining b) mismatch repairc) nucleotide-excision repaird) base-excision repair e) homology-directed repair
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.
Does one have a practical idea of the role of CRISPR/Cas9 in genome editing via cutting of DNA?