CRISPR techniques allow scientists to modify specific genes while sparing all others, thus clarifying the association between a given gene and its consequence to the organism.  If this technology can change the future of Medicine, what specific benefits CRISPR can bring to genetic testing or analysis?  How can CRISPR help to enhance gene therapy or treatment of genetic diseases?

A patient is diagnosed with lung cancer using imaging. To establish the best treatment option, doctors ask to profile the gene expression of the tumor, using a biopsy and microarray analysis. Describe the steps involved in the analysis in detail. The analysis reveals that a new gene therapy could be beneficial to the patient. Explain briefly how gene therapy could be used to cure cancer and detail how the treatment can be formulated for delivery to the patient. To prepare DNA for the gene therapy formulation, it is often important to measure the size of the molecule. Explain how this measurement can be done using agarose gel electrophoresis, taking care to detail the property of the DNA molecule at the basis of the technique. To generate large amounts of DNA to manufacture gene therapy payloads or to be able to see them on gel electrophoresis, specific sequences can be amplified by PCR. For a sequence of a 100 base pairs, calculate the number cycles of PCR required to generate 1 ng of…

CRISPR techniques allow scientists to modify specific genes while sparing all others, thus clarifying the association between a given gene and its consequence to the organism. How can this help to create better antibiotics or to make white blood cells resistant to HIV infection? Give your examples. What is the future of CRISPR?

Briefly explain how a gene can be isolated through positional cloning.

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? mutagenesis with radiation random mutagenesis CRISPR-Cas9 genome editing followed by nonhomologous end joining (NHEJ) CRISPR-Cas9 genome editing followed by homologous recombination (HR) with a donor template oligonucleotide-directed mutagenesis