Thalassemia Essay

Polymerase Chain Reaction was run and the results were placed into an electrophoresis gel to visualize the PCR product. To determine the alleles in the gene, a digest enzyme, Fnu4HI, was placed into the DNA. Tasters would view a cleavage formed at a restriction site in at least one of the alleles, whereas non-tasters would not.

To determine the solo-LTR region I first ran an electronic PCR on s240c3, s165c5 and s399c8 primers. I then aligned the PCR products to each other using a series BLAST searches, identifying a related sequence of ~968bp expected to be the same in each PCR product. Using BLAST I identified the start and end points of this sequence in relation to each of the PCR products of the 3 primers used in the experiment, allowing me to determine the direct repeats and thus pick out the Solo-LTR sequence and pre-integration site. From the size of the solo-LTR sequences, the size of the whole sequence and the identification of the direct repeat sequences I was able to work out the size of the preintegration site by taking the whole PCR product

Method: There is a method of genome editing called CRISPR Cas9. Borrowed from the idea of a genome editing system in bacteria, the Cas-9 protein can snip out a mutated gene and replace it with a normal gene. This will fix the gene making the proteins that it produces go back to normal so it can do its job. When the patient has an offspring, the offspring won’t be affected since the patient now has the normal gene.

Figure 1 Gel Electrophoresis for Replication Taster PTC. The gel is composed of an ethidium bromide stained 3% agarose gel demonstrating DNA fragments which were a depiction of PCR amplification. The agarose gel contains nine loading samples, including from left to right, the MW marker lane 1 precision mol mass standard, lane 2 TB undigested PTC (5µl of DNA, 5µl of master mix P, and 2.5µl of loading dye), lane 3 TB digested PTC (5µl of DNA, 5µl of master mix P, 2µl Fnu4HI, and 3µl of loading dye), lane 4 TB A(L)DH G (10µl DNA, 10µl master mix G, and 5µl loading dye), lane 5 TB A(L)DH A (10µl DNA, 10µl master mix A, and 5µl loading dye), lane 6 MG undigested PTC (5µl of DNA, 5µl of master mix P, and 2.5µl of loading dye), lane 7 MG digested PTC (5µl of DNA, 5µl of master mix P, 2µl Fnu4HI, and 3µl of loading dye), lane 8 MG A(L)DH G (10µl DNA, 10µl master mix G, and 5µl loading dye), lane 9 MG A(L)DH A (10µl DNA, 10µl master mix A, and 5µl loading dye).

Primers dimers occur frequently at a large threshold cycle number ( more than 35 cycles)

Designer babies are babies whose DNA has been modified to include and exclude certain traits. One such example is CRISPR-Cas9, a new technology, giving scientists the ability to control the genetic structure of human cells and use to create designer babies (Sas & Lawrenz, 2017). Additionally, what makes this gene editing attractive is the ability to locate, modify, or even remove deleterious gene diseases (Otieno, 2015). Gene editing might be beneficial to bring a child into this world without extra challenges such as deformities, genetic defects, and illnesses yet, there are serious concerns over this practice. Ultimately, not only is gene editing both difficult and expensive to achieve, but it is also an unreliable way to change natural order

The final, but perhaps most tremendous, genetic controversy is known simply as the “Designer Baby Controversy.” According to Welch and CBS News, scientists Junjiu Huang and his colleagues at Sun Yat-Sen University in Guangzhou, China, “have successfully edited DNA in human embryos, a highly controversial move governing much criticism in the scientific community.” Huang and his team have spliced out a gene called HBB, which causes beta-thalassemia, a fatal blood disease, and successfully replaced it in the embryos with a healthy copy of the gene. This method could be used to fix many other single-gene diseases, and, eventually, modified to remove other multiple-gene disorders, as well. However, two large problems arise from this. The first

Thanks to advancements in gene editing technology, a future with far less genetic defects and diseases is not that far away. Professor Julian Savulescu states that “Gene editing could be used to cure genetic diseases such as cystic fibrosis or thalassaemia (the blood disease that the Chinese researchers were working to eliminate). At present, there are no cures for such diseases.” While disputants will insist that prenatal screening for genetic selection could allow couples to simply choose a healthy embryo, “genetic selection doesn’t benefit patients - it’s not a cure. It merely brings a different person, who is free from disease, into existence. Future

According to data provided in class, the smallest band of Hind III shown on the curve at 2.02 kbp should have traveled 24 cm. As shown in table a, the band with 2.02 kilo base pairs traveled 20.2 cm. III fragment that contained 23.1 kbp traveled only 7.30mm when it should have traveled 11.0 mm. However, the fragment that had 4.36 kbp should have traveled 18mm when it traveled 19mm. This shows the enzymes did not cut right where they were supposed to, resulting from not allowing the fragments to run long enough. An extra half hour or so would have allowed enough time for the fragments to be cut in the right places. After generating a standard curve shown above in example c, the EcoR I fragment sizes were estimated. The largest fragment traveled only 6.79mm, and it was estimated that the fragment was 23.8 kilo base pairs in size. The actual fragment size derived from the provided restriction map was 21.2 kilo base pairs in size. The second smallest fragment traveled 11.6 mm and it was estimated that the fragment was 6.01 kilo base pairs in length. The actual fragment was 5.6kpb in size. The majority of the estimated EcoR I fragments were larger than their actual fragments. This was the result of the Hind III fragments not reaching their end distances in the gel. Since the standard curve was made from the partially traveled Hind III fragments, the estimated numbers of EcoR I were off. The creation of a standard curve was a good way to estimate the DNA fragments sizes, because both the sets of DNA were run at the same time for the same length of time. If one does not have adequate time to travel as far as it should, neither will the other sample being run. EcoR I produced fewer bands than predicted by its restriction map because the smallest band may contain too little DNA to be seen without using a more sensitive dye to illuminate the

The US and Korean team created 58 human embryos while using the CRISPR-Cas9 they were able to repair faulty genes up to 70%. (…)

This beta-globin gene disorder presents an ideal candidate to provide gene edition or correction as a form of therapy. Bearing this in mind, globin gene transfer into a patients own hematopoietic stem cells developing into red blood cells with normal hemoglobin content would be the goal. Transduced hematopoietic stem cells can maintain a long-term clinical effect through a competent erythropoiesis produced by a normal globin chain synthesis ratio. The ultimate goal of HSCs and gene therapy in thalassemia patients is to attain independence from the transfusion therapies without the risks of an ideal matched HSC donor

New gene editing tools/ technologies have made scientist excited about what to come. Researchers say that these tools that can fix human diseases are only a few years away. Chinese researchers have started to edit human embryos, those obtained from local fertility clinic because they contain an extra set of chromosomes. This article is important to my presentation because it not only shows progress from other countries but the enthusiasm for gene editing.

The first is a transition point mutation, in which at nucleotide 845 a guanine in replaced by adenine. This results in a missense mutation that replaces a tyrosine for a cysteine at amino acid 282 (C282Y). This mutation affects the function and structure of the HFE protein; “it prevents the formation of a disulfide bridge in the HFE α3 domain, impairing the β2-microglobulin and the transferrin receptor-1 (TfR1) interactions.”7 TfR1 is required for the uptake of transferrin-bound iron into cells, and β2-microglobulin helps regulate the expression of hepcidin in the liver which is mentioned later in the

The processing included utilising gene therapy to correct the SNP. As mice infected with humanised SCA live for only around 18 months, the untreated mice, which had mutations in both genes, all died; those that had the injections all survived. The results of this study showed that the injected mice had decreased symptoms of SCA, proportionate to that of the control mice, which had only one mutated gene. A second study employed the use of two transgenic mice models and found that, although one model worked markedly better, both gene therapies decreased the level of Hb S and increased Hb level. The researchers altered a βA-globin gene, via an SNP, and placed it within lentiviruses to administer it to the test mice. This led to a general decrease in the number of sickled RBCs within the treated mice. The results from this study also showed that the treated mice had levels of Hb equal to those of

An important step in detecting the single nucleotide changes within the HFE gene that causes hemochromatosis is to first use the process of Polymerase Chain Reaction (PCR) amplification to amplify specific parts of exons 2 and 4. In the PCR amplification specific primers were used independently for each exon 2 and