9. The CRISPR associated protein Cas9 is now being used as a molecular tool for genomeeditingC. In a cell, what happens to a gene that has been cut by a genome editing system? It isrepaired using either non-homologous end joining (NHEJ) or homology dependentrepair (HDR)D. The Cas9 protein requires an RNA molecule to carry out genome editing. Whatsequence requirements does this RNA have?E. Name two limitations of CRISPR/Cas9 genome editing.

C) CRISPR is a tool of genome editing by using the endonuclease cas9, when the gene is cut by genome…

Answered: e CRISPR associated protein tinal

Biology: The Dynamic Science (MindTap Course List)

4th Edition

ISBN:9781305389892

Author:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Publisher:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Chapter18: Dna Technologies: Making And Using Genetically Altered Organisms, And Other Applications

Section: Chapter Questions

Problem 1TYK

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1. What is the function of the CRISPR/ Cas system 2. Briefly outline the components of the CRISPR/Cas system
In 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.
Why are antibiotic resistance markers such as ampR important components of bacterial plasmid cloning vectors? a. The plasmid must have resistance to accept DNA inserts. b. They allow the detection of plasmids that contain an inserted DNA fragment. c. They ensure the presence of the ori site. d. They ensure that the plasmid can be cut by a restriction enzyme. e. They allow identification of bacteria that have taken up a plasmid.
A cDNA and a cloned fragment of genomic DNA share sequences from a mouse gene. What differences do you expect to see between the cDNA and genomic DNA sequences? a. None; they should be identical. b. The genomic DNA might have an intron or introns. c. The genomic DNA might have promoter sequences. d. The genomic DNA might have a poly(A) tail. e. Both b and c are correct.
1. Examine the “RNA-Seq Coverage” and the “FlyBase Genes” tracks in the Genome Browser from left to right. At approximately which coordinate (base position) does the RNA-Seq data start? Remember that you can use the navigation controls at the top of the page to zoom in to the region of interest.
1. Explain how the SNP changes the recognition sequence for the restriction enzyme and affects its ability to cut the DNA. Indicate which of the 2 alleles (taster or non-taster) is cut by the restriction enzyme. 2. Predict the size of the expected DNA fragments after restriction digest for the two alternative SNP sequences (the taster and the non-taster).i.) Taster allele after being cut ii.) Non-taster allele after being cut
1. Why would multi-gene families complicate things in terms of being sure of which member of the family was responsible for a particular phenotype? 2. How could multi-genes families complicate things in terms of using CRISPR to knock out a target gene and achieve a target phenotype?
1. Given the following restriction endonucleases and the sequences of their corresponding restriction sites, which would be LEAST useful for cutting the plasmid and the foreign DNA to be inserted? [The arrows indicate where cuts are made.] (see img) a. EcoRI b. HindIII c. HaeIII d. BamHI 2. EcoRI and HindIII are two different restriction enzymes. If the DNA from two different sources were cut with either EcoRI or HindIII, which of the cut DNA fragments would NOT join together easily so that they could be sealed with ligase? a. E. coli DNA cut with EcoRI and human DNA cut with HindIII b. E. coli DNA cut with HindIII and mouse DNA cut with HindIII c. human DNA cut with EcoRI and chimpanzee DNA cut with EcoRI d. mouse liver DNA cut with HindIII and mouse kidney DNA cut with HindIII
9)A scientist compares the sequence of a disease gene and a healthy gene and finds that these two genes are identical except for one G in place of a T at position 256 in the gene. This type of mutation is a(n): Select one: a. deletion b. insertion c. silent mutation d. frameshift e. substitution mutation
1. What makes CRISP particularly useful compared to other genetic engineering techniques? A. CRISPR can add DNA to multiple cells at the same time B. CRISPR works better in bacteria C. CRISPR can target one specific area of DNA to cut D. CRISP is capable of transporting DNA through the cell walls of plants 2. MOST viruses are capable of infecting A. many different species B. many cell types in the same species C. only specific cell types that have a particular receptor
1. Which of the following explains why prokaryotes have a bigger gene-to-genome size ratio compared to eukaryotes? A. Prokaryotic genome contains multiple promoters and repressors that control of a single gene.B. Prokaryotic genome is completely devoid of non-coding sequences.C. Prokaryotic genome contains multiple genes that are under the control of a common promoterand repressor.D. Prokaryotic genome contains one promoter region for each gene. 2. Which of the following can explain why eukaryotes have smaller gene-to-genome size ratio compared to prokaryotes?A. Genome size do not correlate with complexity in eukaryotes.B. The percentage of genes is inversely correlated with genome size in eukaryotes.C. Eukaryotic genome contains a lot of coding sequences.D. Eukaryotic genome contains a lot of non-coding sequences.
2) The authors investigate if the switch from the polymerase to the exonuclease site involves releasing and re-binding of the DNA or if it utilizes an intramolecular rearrangement. To accomplish this, they use a primer-extension assay. How is a primer extension assay performed? In answering this question, be sure to mention a.) why heparin is used, b.) how they generate mismatched primers, and c.) how they visualize only the primer strand and not the template strand on their gel.

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