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Drafts
Somatic Gene Cell Editing
Somatic cell gene editing modifies a subjects DNA in order to try and treat a disease caused by a genetic mutation in a specific type of cell. Only the targeted cell type will be affected through somatic gene editing. One way to perform this is the use of CRISPR techniques on blood stem cells to try and fix the genetic mutation. This will change the subject’s blood cells, but will not have any impact on their sperm or eggs. This means that the edited gene will not be passed down to future generations.
Data Analysis- Proposal
All this information should be recorded in a chart that displays the measurements of the volunteer and host species in individual pots and group pots. Groups should look for similarities and differences between the volunteer species present in each plant. They should also look for differences in growth between the two host plants. If the greenhouses can provide information about the habitat these plants are grown in, like soil and climate information, groups should use this information to discuss possible reasons for their differences. Likewise, it may be beneficial for groups to attain information on how old each host plant is. Groups should apply these similarities and differences in an organized discussion section. This will require additional research to be done on the specific volunteer and host species and their ideal growth environments.
Gene Analysis Results 1
We next narrowed our literature search in pursuit of gene function of our specific gene. A search of Bradi1g25180 did not yield any results, so we switched our focus to homologous proteins. Bradi1g25180 was searched on www.arabidopsis.org in order to find homologous proteins. There were twenty homologs identified, but searches for all but one yielded no papers about specific function. The arabidopsis thaliana gene AT4g21390 (gene B120), also known as T6K22.120 according to Uniprot, is homologous to Bradi1g25180 according to www.arabidopsis.org. A search for “AT4g21390” yielded nothing, but a search for “T6K22.120” yielded a paper in which the gene was hypothesized to be the Arabidopsis thaliana counterpart to the S-locus receptor kinase (SRK) when compared to SRK in Arabidopsis lyrata (Kusaba et al. 2001). However, the authors ultimately concluded that the homologous counterpart was actually the adjacent ARK3 gene, at position 110, and that gene B120 is related to the BRLK gene in Brassica oleracea.
Root Growth and Root Respiration
The relationship between root growth rate and root respiration is said to be by maintaining low root respiration rates (low maintenance cost) high-temperature adaptation and root survival is possible for many species of plants. The Q10 ratio is the temperature sensitivity of respiration. It is essentially the ratio of respiration rates per 10 degree Celsius increase in temperature. Long term exposure to temperatures can result in respiratory acclimation which is essentially the adjustment of respiration rates to compensate for temperature change. There is not a lot of data on root acclimation at higher temperatures and this may suggest that the root respiration acclimation varies between plant species.
bio 559 local hero part3
To test if PLCζ was the sole calcium trigger in sperm, Fissore’s team created a PLCζ-null mouse using CRISPR/Cas9. Using CRISPR, they targeted the Plcz1 gene in fertilized mouse eggs. They came up with two strategies as to go about this: Cas9WT with a single sgRNA and Cas9D10A nickase with paired sgRNAs. A line of sperm where Cas9D10A induced a 22 nucleotide deletion in the target area (Plcz1em1Jparr), and another where Cas9WT induced a 17 nucleotide deletion in another area (Plczl em2Jparr) were created. These deletions created two mutant alleles for the Plcz1 gene. When injected into mouse eggs, the mutant cRNA did not induce calcium oscillations confirming that the mutants were PLCζ-null (Plcz1-/-).
They found that the loss of PLCζ did not affect the sperms quality or its ability to bind and fuse with an egg. Wild-type sperm (Plcz1+) and Plcz1-/- sperm (Plcz1-) response to progesterone or ionoycin were undistinguishable from each other. When testing the Plcz1+ sperm in wild-type mouse eggs, the sperm induced calcium oscillations “symptomatic of those seen at fertilization in nearly all eggs.” The Plcz1- did not. This was then confirmed with using in vitro fertilization of zone pellucid-free eggs (eggs without a glycoprotein layer around its plasma membrane). 16 out of 34 eggs with Plcz1+ sperm showed Ca2+ oscillations and none of the eggs with Plcz1 em1Jparr derived sperm show any oscillations. His team showed the first direct evidence that PLCζ is the “sole physiological trigger of the Ca2+ oscillations responsible for egg activation in mammals.”
Bio 559 local hero part2
On campus, his lab focus is a continuation of his Ph.D. work with in-vitro fertilization in mammal cells and how exactly sperm begins a calcium released oscillations in the egg. On this topic, he was a part of a paper published in 2017. It is titled PLCζ is the physiological trigger of the Ca2+ oscillations that induced embryogenesis in mammals but conception can occur in its absence. The goal of the study being how exactly sperm triggers calcium oscillations in the egg. This study is important for humans as infertility is a health problem that affects about 1 in 7 couples. The paper stated that in other studies there was evidence that a protein called phospholipase C zeta (PLCζ) trigger calcium release in sperm.
Local Hero Paper for bio 559
Rafael A. Fissore is a professor and department head at the University of Massachusetts and has been a member of the community for decades. He graduated with a Ph.D. from the university in 1993 in Animal Science. By that time, he had already been a part of a published paper on calcium concentration in bovine (cattle) eggs in 1992. He continued his work with postdoctoral training at Harvard Medical School. The focus there being chemical signals to induce reproduction in animals, one of the signals being calcium. In 2002 he received the CFNR Outstanding Research Award. In 2004, as an associate professor of Animal Science at the University of Massachusetts Amherst, he gave a talk on his fertilization research at the Activated Egg Symposium. He has also filed three patents between early 2003 and mid 2008, by himself and with teams, all towards his field of research in mammalian fertility.
Fissore is currently the department head of the Department of Veterinary and Animal Sciences on campus. In 2008, he received a Distinguished Faculty Lecturer honor and the Chancellor’s Medal. He also teaches the honors Animal Science 521: Physiology of Reproduction class. The class focuses on recent studies on cellular and molecular aspects on mammalian fertilization. It also looks at technical and ethical problems when applying new technologies for assisted reproduction and cloning, and how it applies to agriculture, humans, and the natural world.
Irisin and Alzheimer’s Prevention
Exercise assists in maintaining the health of a person. New studies have found that exercise can also assist the brain in resisting Alzheimer’s disease. Mice were used in the study to see whether irisin had a role in increasing brain function. Mice with dementia were injected with the irisin protein and was found to do better on memory tests. Irisin is created through the cleavage of FNDC5 protein. FNDC5 increases through exercising, when you exercise more it would cause more FNDC5 to be cleaved and more irisin secretion. Irisin works by causing a change in gene expression of the beta amyloid.
Gap Junction Inhibitors
Another therapy approach described to target the survival function of the metastatic niche are gap junction inhibitors. In the brain metastatic niche, astrocytes were shown to interact with disseminated tumor cells (DTCs) through gap junctions in order to further their survival through the second messenger cGAMP and the activation of the STING pathway as well as interferon-alpha and tumor necrosis factor (Chen et al. 2016). Therefore, by inhibiting these gap junctions, cancerous cells are not able to utilize surrounding cells to further their survival. Some of these gap junction inhibitors include the FDA-approved tonaberset and meclofenamate. This therapy is useful in halting cell survival but is systemic as many cells throughout the body interact through gap junctions, including those that are healthy. Instead, these gap junction inhibitors may be better utilized in synergy with a more specific treatment to target only cancerous cells.
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