The goal of most gene therapies is to insert a healthy copy of a gene into the genome. Besides CRISPR, techniques for integrating DNA into the genome do not target that DNA to a particular genomic location and thus different cells end up with the DNA sequence in different positions. A. It has been shown that where the healthy gene integrates is heavily influenced by chromatin. Why?
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The goal of most gene therapies is to insert a healthy copy of a gene into the genome. Besides CRISPR, techniques for integrating DNA into the genome do not target that DNA to a particular genomic location and thus different cells end up with the DNA sequence in different positions.
A. It has been shown that where the healthy gene integrates is heavily influenced by chromatin. Why?
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- The goal of most gene therapies is to insert a healthy copy of a gene into the genome. Besides CRISPR, techniques for integrating DNA into the genome do not target that DNA to a particular genomic location and thus different cells end up with the DNA sequence in different positions. A. It has been shown that where the healthy gene integrates is heavily influenced by chromatin. Why? B. The same gene integrated into different locations in the genome will be expressed differently. Why? C. Over time the expression of an integrated gene will change, usually decreasing. Describe how you think the cell downregulates expression of an integrated gene.The goal of most gene therapies is to insert a healthy copy of a gene into the genome. Besides CRISPR, techniques for integrating DNA into the genome do not target that DNA to a particular genomic location and thus different cells end up with the DNA sequence in different positions. B. The same gene integrated into different locations in the genome will be expressed differently. Why?Describe the main technique for amplifying a segment of DNA (like the one you suspect is involved in Lee’s cancer) from a complex mixture of genomic DNA. Remember that the entire human genome sequence is known. (Hint: This is a technique that is commonly used by laboratories that do genetic testing and various other applications of molecular biology.)
- The technique of fluorescence in situ hybridization (FISH) is described. This is another method for examining sequence complexity within a genome. In this method, a DNA sequence, such as a particular gene sequence, can be detected within an intact chromosome by using a DNA probe that is complementary to the sequence.For example, let’s consider the β-globin gene, which isfound on human chromosome 11. A probe complementary to theβ-globin gene binds to that gene and shows up as a brightly colored spot on human chromosome 11. In this way, researchers can detectwhere the β-globin gene is located within a set of chromosomes. Becausethe β-globin gene is unique and because human cells are diploid(i.e., have two copies of each chromosome), a FISH experimentshows two bright spots per cell; the probe binds to each copy ofchromosome 11. What would you expect to see if you used thefollowing types of probes?A. A probe complementary to the Alu sequenceB. A probe complementary to a tandem array near…From your knowledge about DNA microarray, answer the following: If the expression microarray experiment was done with a normal sample and a suspected sample, after reading the color pattern resulted from the experiment it was recorded that “gene A22” is expressed in the suspected sample. The gene A22 is clinically linked to colon cancer. Answer the following: What is the expected color of the spot on the microarray which represents this gene? What is your interpretation of the suspected sample; is it a cancer sample or not and explain why?#3) Ligase catalyzes a reaction between the 5' phosphate and the 3' hydroxyl groups at the end of DNA molecules. The enzyme calf intestinal phosphatase catalyzes the removal of the 5' phosphate from DNA molecules. What would be the consequence of treating a cloning vector, before ligation, with calf intestinal phosphatase?
- Genes with highly similar sequence are often located adjacent one another in the genome. Gene duplication commonly arises from errors in replication. When the organization of such adjacent genes is in an inverted orientation, this can reduce the expression of other genes that have similar sequence and are located on other chromosomes. Explain the mechanism of how this generally occurs. i need a detailed explanation, please. I need to know what the mechanism is. How it happens and why. Please. ThanksWith the use of well-illustrated diagrams, reconstruct the entire cloning process by explaining different stages of the cloning process that involves the following: d. Screening/selection of hosts containing the intended recombinant plasmid. For this stage(d), discuss the importance of a second marker that can be used for screening of genomic DNA for colonies containing the pka-1 under the principle of insertional inactivation. This should be properly explained using all the attributes of the plasmid described above.In a clinical context, doctors are evaluating a therapy for a new patient (say patient B) that they have reasons to believe might develop a cancer similar to another patient they treated successfully (patient A). They know that the severity of the cancer is mainly associated with mutations in a specific gene (BRCAI). Suggest a technique that can be used to rapidly assess the similarity between the genetic panerns of patients A and B, without the need to sequence the entire gene, and briefly describe it. A team of scientists are interested in the amplification of a specific DNA fragment in a large plasmid of about 10000 bps. (0) (11) The sequence the scientists are interested in is 5'-CATTGATTATTG[...JATCAATTACGGG-3" 3-GTAACTAATAAG[...]TAGTTAATGCCC-5* Where [...] indicates a longer 100bps sequence. Provide two possible primers that the scientists should use to address their need, if they want to be sure they specifically address this region in the entire plasmid. Briefly explain the…
- DNA hypermethylation (an excess of methylation) is associated with many neurological disorders, including a potential role in Alzheimer's disease. a. When comparing individuals with and without Alzheimer's, which of the following 'omics techniques (exome sequencing, whole genome sequencing, transcriptomics, or proteomics) would you expect to be the most informative if your goal is to locate potentially causative methylation differences? Briefly justify your answer. b. The pdCas9-Tet1-CD enzyme (Xu et al Cell Discov. 2016) fuses an enzyme that can carry out cytosine demethylation to a mutant version of the CRISPR/Cas9 enzyme that localizes to DNA using a guide RNA in exactly the same manner as we discussed in class for standard Cas9, except that this version does not cut the genome. This demethylase enzyme can then act to remove DNA methylation proximal to where it is stably bound. Imagine that you have identified a gene that is hypermethylated specifically in patients with Alzheimer's…Northern blotting, RT-PCR, and microarrays can be used to analyze gene expression. A lab studies yeast cells, comparing their growth in two different sugars, glucose and galactose. One student is comparing expression of the gene HMG2 under these two conditions. Which technique(s) could he use and why? Another student wants to compare expression of all the genes on chromosome 4, of which there are approximately 800. What technique(s) could she use and why?Genome annotation refers to ... 1.) lining up overlapping regions in short shotgun sequencing reads to assemble larger contiguous DNA sequences (contigs/scaffolds). 2.) using long-read sequencing platforms (such as PacBio or Oxford Nanopore) to gather information about the epigenetic status of each region of a sequenced genome. 3.) the process of predicting which parts of a genome sequence code for functional products (such as protein-coding genes), what those products do, and assigning them names. 4.) sequencing messengeRNA measure the relative expression levels of genes in one or more tissue samples.