You would like to create a mouse model of the human disease Huntington's chorea, a dominant disease caused by a gain of function mutation in the human HD gene. Which of the following strategies would be suitable? Select all that apply 0 Use CRISPR-Cas9 to introduce frameshift mutations in the mouse HD gene; breed the resulting mutant mice to generate homozygous mutants 0 Introduce the wild type human HD allele as a transgene into mouse genome; select transgenics where human HD replaced the wild type mouse HD gene via homologous recombination Introduce the mutant human HD allele as a transgene into the mouse genome with transgene integration anywhere in the mouse genome Use CRISPR-Cas9 technology to cut the mouse HD gene; supply the mutant human HD allele as a repair template and select for transgenics where the the mutant human HD replaced the mouse HD gene

You would like to create a mouse model of the human disease Huntington's chorea, a dominant disease caused by a gain of function mutation in the human HD gene. Which of the following strategies would be suitable? Select all that apply 0 Use CRISPR-Cas9 to introduce frameshift mutations in the mouse HD gene; breed the resulting mutant mice to generate homozygous mutants 0 Introduce the wild type human HD allele as a transgene into mouse genome; select transgenics where human HD replaced the wild type mouse HD gene via homologous recombination Introduce the mutant human HD allele as a transgene into the mouse genome with transgene integration anywhere in the mouse genome Use CRISPR-Cas9 technology to cut the mouse HD gene; supply the mutant human HD allele as a repair template and select for transgenics where the the mutant human HD replaced the mouse HD gene

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter13: An Introduction To Genetic Technology

Section: Chapter Questions

Problem 20QP: Analyzing Cloned Sequences A base change (A to T) is the mutational event that created the mutant...

See similar textbooks

Similar questions

Gerald Rubin and Allan Spradling devised a method of introducinga transposon into Drosophila. This approach has been importantfor the transposon tagging of many Drosophila genes. Theresearchers began with a P element that had been cloned on a plasmid.Using cloning methods, they inserted the wild-type allele for the rosygene into the P element in this plasmid. The recessive allele, rosy,results in a rosy eye color, while the wild-type allele, rosy+, producesred eyes. The plasmid also had an intact transposase gene.The cloned DNA is shown to the right.Rubin and Spradling used a micropipette to inject this DNAinto regions of embryos that would later become reproductivecells. These embryos were originally homozygous for the recessiverosy allele. However, the P element carrying the rosy+ allele could “hop” out of the plasmid and into a chromosome of the cells that were destined to become germ cells (i.e., sperm or egg cells). After the embryos had matured to adults, the flies were then…
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.
What would three possible reasons be that a PCR wouldn't work after cloning a TAQ sequence into a His-tag vector making three different constructs and verifying via SDS-PAGE that the TAQ protein is there?
The sequence at one end of one strand of the Drosophilatransposon Mariner is shown below (dots indicatesequences within the transposon):5′ TTAGTTTGGCAAATATCTCCCTTCCGCCTTTTTGATCTTATGT... 3′You obtain a mutant bacterial strain tagged with anengineered Mariner transposon, cut the genomicDNA from this strain with the restriction enzymeMboI (whose recognition site is ^GATC), and circularize the resultant DNA fragments by diluting therestriction enzyme digest and adding DNA ligase.a. Design two 17 bp PCR primers that you could useto identify (by inverse PCR) the gene into whichthe transposon inserted.b. What DNA sequence will be amplified from thecircularized fragments of the mutant genome?Show the extent of this DNA sequence on a mapof the genome of the mutant strain, indicating thelocations of the transposon insertion and any relevant sites for the enzyme MboI.
You know from your calculationsthat only a small proportion of the human genome is represented, even when the entire class results areconsidered. Therefore, the chance of finding a particular single-copy gene in your library is very small.Outline a strategy for constructing a genomic DNA library more representative of the entire humangenome. You will need to consider alternative vectors and the efficiency of transformation of thebacterial cells.
I get everything else in this problem other than the third option: Introduce the mutant human HD allele as a transgene into the mouse genome with transgene integration anywhere in the mouse genome. Why is the first question (left) okay with introducing mutant human HD allele and the second question (right) is not? I heard that introducing allele without using CRISPR-Cas9 is very rare and difficult. If so, how does it work in the first problem (Hungtinton's chorea)?
The ability to selectively modify the genome in the mouse has revolutionized mouse genetics. Outline the procedure for generating a knockout mouse at a specific genetic locus. How can the loxP-Cre system be used to conditionally knock out a gene? What is an important medical application of knockout mice?
You are trying to study the effects of Drug A on the expression of Gene X in a tumor sample (lane 3). You perform an RT- PCR reaction using a set of prímers to amplify a region of Gene X that is about 600bp in length. The forward and reverse primers are on 2 separate exons. You run your target CDNA fragment on an agarose gel (lane 3). Lanes 1 and 2 are hypothetical control lanes to test for 9DNA contamination. 1 3 1000 bp 600 bp Based on the gel image, which of the following statements is correct? 1. The samples loaded in lanes 1 and 2 were not reverse transcribed prior to PCR. 2. Results from lane 2 suggest that 9DNA contamination is present. 3. The upper band in lane 2 represents 9DNA. 4. The fact that lane 1 is blank is indicative that Drug A doesn't lead to expression of Gene X. O A. 1, 2, and 3 O B. 1 and 3 O C. 2 and 4 O D. 4 only O E. All of 1, 2, 3 and 4 are correct
You identify a proflavin-generated allele of a genethat produces a 110-amino acid polypeptide ratherthan the usual 157-amino acid protein. After subjectingthis mutant allele to extensive proflavin mutagenesis,you are able to find a number of intragenic suppressorslocated in the part of the gene between the sequencesencoding the N terminus of the protein and the originalmutation, but no suppressors located in the regionbetween the original mutation and the sequencesencoding the usual C terminus of the protein. Whydo you think this is the case?
Mouse models for human genetic diseases are potentially powerful tools to help geneticists understand thecause of the aberrant phenotypes and develop newtherapeutic measures. However, such mice are not always as useful to investigators as it might seem at firstglance. Suppose that you have a mouse knockoutmodel for a human disease caused by homozygosityfor a null allele of a gene. Discuss how the followingsituations might complicate investigations of the human disease based on this mouse model.a. Mice have a shorter life span than humans.b. Mice homozygous for certain knockout mutationsdie in utero.c. Mouse genomes may have additional copies of thegene whose mutation causes the disease in humans.
Mouse models for human genetic diseases are potentially powerful tools to help geneticists understand thecause of the aberrant phenotypes and develop newtherapeutic measures. However, such mice are not always as useful to investigators as it might seem at firstglance. Suppose that you have a mouse knockoutmodel for a human disease caused by homozygosityfor a null allele of a gene. Discuss how the followingsituations might complicate investigations of the human disease based on this mouse model.a. Mice have a shorter life span than humans.b. Mice homozygous for certain knockout mutationsdie in utero.c. Mouse genomes may have additional copies of thegene whose mutation causes the disease in humans.d. Mice from different inbred lines homozygous forthe same gene knockout vary in the penetrance andexpressivity of the phenotype.e. Manipulations to create the knockout mouse, suchas the presence of a drug resistance gene that allowsthe selection of cells containing the knockout (seeFig. 18.9),…
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?