a gm animal that may be approved for human consumption by the time this book is published is a super muscly pig made by inactivation of the myostatin gene. During normal development, myostatin protein prevents the overgrowth of muscles. (Given that effective deletions can be made in the genome by using CRISPR/Cas9. It targets deletions of varied length. By this technique, the transcriptional status of the target gene is not affected. The gene deletions produced by CRISPR/Cas9 lead to the production of the correct type of junctions at very high frequencies), how could the super muscly pig have been generated?Answer the questions in the photo as well. ( i included my answer for 22a in the question above, if you believe you have a better answer by all means include that as well) 35. Recently, scientists have used a mouse model for Duchenne muscular dystrophy (called themdx mouse) to test whether Cas9 and an sgRNA could be an effective therapy for thisdisease. The cause of muscular dystrophy is homozygosity or hemizygosity for loss-of-function mutations in the X-linked Dmd gene. The mdx mouse has a nonsense mutation inexon 23 (of 79 exons) of Dmd. Researchers tested a technique called exon skipping; their ideawas to use CRISPR/Cas9 to delete exon 23 in the mutant Dmd gene in the mdx mouse. AAVvectors with genes that express Cas9 and two different sgRNAs were injected into themuscles of adult mice. In about 10% of the muscle cells, exon 23 was deleted, and functionaldystrophin protein was detected. Some muscle function was restored, although only to a smallextent.a. Draw a diagram of exon 23 and the introns that flank it. On your diagram, draw thelocations where the sgRNAs would hybridize with the genomic DNA. Explain how thedeletion of exon 23 would take place.b. Design a PCR assay to determine if exon 23 is deleted from the genomic DNA of cellclones. Where would the PCR primers hybridize, and how would you be able to tell if theexon was deleted?c. Skipping exon 23 restored dystrophin protein function at least partially. What does thissay about the amino acids encoded by exon 23?d. Considering your answer to part (c), would the exon-skipping strategy work for allDmd point mutations that cause muscular dystrophy?e. What must be true about exons 22, 23, and 24 that would allow the researchers toconsider this exon-skipping strategy? (Hint: See Problems 28 (9 and 29 9.) 22. Scientists now routinely use CRISPR/Cas9 to make defined deletions of a gene that canremove several kb of DNA from the genome. This method is possible even in cells defectivein homologous recombination, as long as the cells can still perform nonhomologous end-joining (NHEJ).a. How could researchers make such deletions?b. A GM animal that may be approved for human consumption by the time this book ispublished is a “super-muscly" pig made by inactivation of the myostatin gene. Page 643During normal development, Myostatin protein prevents the overgrowth ofmuscles. Given your answer to part (a), how could the super-muscly pig have beengenerated?

Hi! Since you posted many questions, we will be answering only the first one for you (question 22).…

Answered: 22. Scientists now routinely use…

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

Related questions

Concept explainers

Question

a gm animal that may be approved for human consumption by the time this book is published is a super muscly pig made by inactivation of the myostatin gene. During normal development, myostatin protein prevents the overgrowth of muscles. (Given that effective deletions can be made in the genome by using CRISPR/Cas9. It targets deletions of varied length. By this technique, the transcriptional status of the target gene is not affected. The gene deletions produced by CRISPR/Cas9 lead to the production of the correct type of junctions at very high frequencies), how could the super muscly pig have been generated?

Answer the questions in the photo as well. ( i included my answer for 22a in the question above, if you believe you have a better answer by all means include that as well)

35. Recently, scientists have used a mouse model for Duchenne muscular dystrophy (called the
mdx mouse) to test whether Cas9 and an sgRNA could be an effective therapy for this
disease. The cause of muscular dystrophy is homozygosity or hemizygosity for loss-of-
function mutations in the X-linked Dmd gene. The mdx mouse has a nonsense mutation in
exon 23 (of 79 exons) of Dmd. Researchers tested a technique called exon skipping; their idea
was to use CRISPR/Cas9 to delete exon 23 in the mutant Dmd gene in the mdx mouse. AAV
vectors with genes that express Cas9 and two different sgRNAs were injected into the
muscles of adult mice. In about 10% of the muscle cells, exon 23 was deleted, and functional
dystrophin protein was detected. Some muscle function was restored, although only to a small
extent.
a. Draw a diagram of exon 23 and the introns that flank it. On your diagram, draw the
locations where the sgRNAs would hybridize with the genomic DNA. Explain how the
deletion of exon 23 would take place.
b. Design a PCR assay to determine if exon 23 is deleted from the genomic DNA of cell
clones. Where would the PCR primers hybridize, and how would you be able to tell if the
exon was deleted?
c. Skipping exon 23 restored dystrophin protein function at least partially. What does this
say about the amino acids encoded by exon 23?
d. Considering your answer to part (c), would the exon-skipping strategy work for all
Dmd point mutations that cause muscular dystrophy?
e. What must be true about exons 22, 23, and 24 that would allow the researchers to
consider this exon-skipping strategy? (Hint: See Problems 28 (9 and 29 9.)

22. Scientists now routinely use CRISPR/Cas9 to make defined deletions of a gene that can
remove several kb of DNA from the genome. This method is possible even in cells defective
in homologous recombination, as long as the cells can still perform nonhomologous end-
joining (NHEJ).
a. How could researchers make such deletions?
b. A GM animal that may be approved for human consumption by the time this book is
published is a “super-muscly" pig made by inactivation of the myostatin gene. Page 643
During normal development, Myostatin protein prevents the overgrowth of
muscles. Given your answer to part (a), how could the super-muscly pig have been
generated?

Expert Solution

Trending now

This is a popular solution!

Step by step

Solved in 3 steps

SEE SOLUTION Check out a sample Q&A here

Knowledge Booster

Learn more about

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.