A scientist wants to make a random mutation in a specific gene to investigate that gene's function. Which of the following methods is she likely to have used to make this change? O mutagenesis with radiation O random mutagenesis CRISPR-Cas9 genome editing followed by nonhomologous end joining (NHEJ) CRISPR-Cas9 template genome editing followed by homologous recombination (HR) with a donor oligonucleotide-directed mutagenesis

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?

In the following "gene library" cloning experiment Digested genomic DNA AmpR gene TCR gene TCR is tetracycline resistant marker, AmpR is ampicillin resistant marker and BamHI is the unique restriction enzyme on plasmid. A PhD student digests/cuts the plasmids with BamHI restriction enzyme and the genomic DNA with EcoRI restriction enzyme. After performing the cloning experiment and obtaining colonies on a selection plate, the obtained cells will be ..... (Hint: this question is even more challenging; the PhD student was later demoted to an MSc student). a) resistant to ampicillin and tetracycline b) sensitive to tetracycline and ampicillin c) resistant to tetracycline and sensitive to ampicillin d) resistant to ampicillin and sensitive to tetracycline e) sensitive to ampicillin and tetracycline BamHI

If a researcher wanted to utilize CRISPR/Cas9 to target a gene of interest in their experimental system (i.e. Drosophila) which of the following would they customize for this purpose Group of answer choices Guiding region of sgRNA None of the above, CRISPR can only function in bacterial cells Cas9 enzyme Scaffold region of sgRNA

What is the purpose of the rat liver extract in this procedure?

63.(2 correct choices)

Answer the following questions about CRISPR below: A.What is a PAM sequence? B.How does the Cas-9/RNA sequence target a piece of DNA for destruction? C.How is the DNA repaired after being cut by CRISPR/Cas-9?

Why do some cells grow on the control plate in the absence of a mutagen?

You are employed in a gene therapy laboratory to test the effectiveness of a vector for correcting the DF508 –CFTR mutation.  The vector uses CRISPR –CAS9 genome editing to correct the mutated gene in situ (CFTR-CRISPR). An Ussing chamber experiment was performed using nasal airway cells obtained from a patient with the DF508 CFTR mutation which were transformed with either a control vector consisting of an inert CRISPR-CAS-9 construct (CONTROL) or the active, gene correcting CRISPR CAS-9 construct (CFTR-CRISPR).  The following transepithelial voltage values were obtained for the drug regimen shown under open circuit and current clamp (injected current of 10mA) conditions: CONTROL   Baseline Amiloride Forskolin Inh172 Vte (mV) -78 -2 -4 -3 DVRt (mV) 8 16 7.5 13 R (W.cm-2) 800 1600 750 1300 I (mA.cm-2) 98 1 5 2 CFTR-CRISPR Vte (mV) -38 -6 -23 -9 DVRt (mV) 9 27 6 11 R (W.cm-2) 900 2700 600…