Drafts

The goal of the research is to validate a CRISPR/CAS9 inactivation treatment for the P23H mutant for Renintitis pigments and put the treatment to test in vivo using the AAV delivery system Experiment 1 that was shown in the study tested whether RHOgene allele-specific targeting could be achieved in vivo. To do this, the researchers attempted to deliver the CRISPER CAS9 VQR system components in the retina of mice by invivo electroporation. To do this, the researchers devised a vector that pCAG vector was electroporated with GFP reporter in the RHO Het eyes.  The invivo efficiency was assessed by harvesting the eyes and purified by FACS sorting and molecular analysis was done.  The mutation frequency of the wt rho gene was also analyzed to test the effect of treatment on Wt cells. From experiment 1, it has been noted that the P23H mutation had high rates of frameshift mutation which leads to the inactivation of RHO allele. The mutant allele is not effective in treating the disease. To asses, this problem The RHO gene mutations that occurred the most often were overexpressed in mice P19 cell line, and colocalization of the Rho n terminus antibody and Rho c terminus antibody.  because the experiment 1 and two resulted in an efficient delivery of CRISPR/CAS9 elements in the retina, with targeting rate of the mutant gene, the researchers attempted to asses the photoreceptor degeneration in mice that was treated through an histological analysis to see what the result of the treatment in the degeneration of the photoreceptor cell. The histological study was done at 1 month.  The same histological study was done again at 3months. To test the injection method of the treatment, the intravitreal injection of AAV2 variant was given, and the histological studies of the retina were performed for the expression of transgenes in the tissue as opposed to the retina specifically. The AAV-PHP mediated targeting of mutation of the whole retina was used with CRISPER/CAS9 was used and the two different approaches are both used, and the efficiency of treatment was assessed for the genetic change. In this experiment, the possibility of enriching for the editing events using GFP transduction using FACS sorting. Using the data that is observed in experiment 6, the researches subjected the High GFP and low GFP cells to NGS analysis to refine the indel analysis. To do this, a PCR of the RHO allele was done to asses their incidence and the editing frequency in retinal samples. This experiment aimed to confirm the selectivity of the approach for the P24H RHO allele.

Cell splitting is the process in which a cell line is moved to a different flask in order to keep the cell line alive. Cell splitting must be done before the cell population gets too large. If the cell population gets too big, the cells will overcrowd the flask they are in and die as a consequence. In the bioimaging lab, a course I am currently taking. In this class, my lab partner and I are working with a line of LLC-Pk1 epithelial cells., and currently have three flasks of cells for our project. When the time comes for splitting, we follow a specific protocol. First, we prepare the new flask that the cells are going into with medium and correct labeling. We must then remove the medium that is already in the flasks. After that we wash the cells with about 1 mL of warm PBS (phosphate-buffered saline) and after rocking the flask and making sure the PBS has washed thoroughly, the PBS is removed and cells that have died have been removed from the flask. The next step is to add about 0.5 mL of trypsin, and then incubate the flask for about 3 minutes at 37 degrees Celsius. This process removes cells from the bottom of the flask so that they can now be transferred to the next flask. The trypsin reaction is stopped b adding about 1 mL of cold medium to the flask and triturating thoroughly Finally, we can now put 5 or so drops from the old flask into the new flask, where the cells now have more room to grow until they need to be split again.

Domestic dogs also have “the K locus, whose genetic characteristics affect the melanocortin pathway.” (Candille et al., 2007) The β-Defensin gene produces a ligand (named β-Defensin) that competitively binds to MC1R. DefB103 or K alleles are discussed in this paper (K^B > k^y). Although K^B is a mutant allele, it is dominant because it has a higher affinity for MC1R than k^y. According to a diagram by Candille et al. (2007), in the presence of Agouti and functional MC1R, there is a stronger affinity for the K^B allele to bind to the receptor. When bound to MC1R, β-Defensin induces synthesis of eumelanin.

Seeing in our full range of colors is something a lot people might take for granted daily. A common alternative is a condition called color-blindness, the most common type being red-green color blindness. Color blindness occurs because the color photoreceptors in our eyes, known as cones, have a deficiency in responding to the proper wavelengths of light. Color blindness is therefore not actually a type of blindness, just a deficiency in perceiving color. In red-green color blindness, the affected individual has difficulty distinguishing between red and green, primarily, but color blindness often affects the whole visible color spectrum. This condition is an X-linked recessive disorder, which means that males are more easily affected by this than females. This is because males have one X chromosome and one Y chromosome, and therefore only need one recessive X  chromosome from the mother to have this condition. This is different from females that need two copies of the recessive X chromosome, one from each parent. Multiple companies now sell glasses that can correct for the wearer's color blindness. This is the best fix we have currently, as there are no surgical procedures or drugs to take that can help curb the condition.

Different partitioning genes (encoding Par proteins) allowed the zygote to part into two poles. Par proteins turned out to be homologous in most eukaryotes (including humans). Loss of posterior PARs lead to expansion of anterior and vice versa. This all suggested a competition of some sort between the different PAR proteins. Kinase domains in the proteins anterior Par kinase domains phosphorylated the lipid binding domains of posterior Par proteins to stop them from accessing the membrane vice versa.

Recently in my lab I have been segmenting light sheet microscopy images of the entire slug brain. The animal used was an adult sea slug with a fully developed nervous system. Sea slugs are interesting neural subjects because their digestive system extends right through the brain. The adult slug brain contains 2 rhinophore ganglia, 2 cerebral pleural ganglia, 2 pedal ganglia, and 2 buccal ganglia. I have also used the software to identify the eye in these images, the optic lobe, and the statocyst, a small sensory organs that aids in balance and orientation. The different ganglion lobes are connected to each other via commisures, although these are harder to see than the bright ganglia in the light sheet images. 

In addition to brain segemntation, I am also completing behavioral hormone experiments in juvenile sea slugs. The hormones in question have already been tested in adult slugs, so hopefully the results of my experiments can give some insight about sensory organ development in this species. 

This poster, titled “Permanent Chemotherapy Induced Alopecia in Young Breast Cancer Patients” does not have an attractive design. White space and alignment is good but the coloring is poor and does not catch your eye. The font and typography is consistent and clean throughout the poster. The layout of the poster is appropriate, except for the lack of an abstract. I do not think posters usually have abstracts, and that the conclusion usually just summarizes everything. Each section is focused and clear with the direct message presented appropriately.

            The writing is presented in bullet points, something that makes the main information easier to catch but looks a little unprofessional. There are no typos on this poster, and overall it seems correct and informative about the findings of alopecia in breast cancer patients. I think the only things I might change about it are the coloring and possibly adding one more figure on the left side. 

Unfortunately, without government intervention and funding the widespread implementation of seaweed aquaculture cannot provide its benefits towards climate change mitigation. Despite all its benefits, seaweed aquaculture is not a ‘silver bullet’ for carbon sequestering. Overall, macroalgae do not have a large impact on the increasing carbon problem the world is facing. The issue is as mentioned, that governments have yet to realize this resource is at their fingertips. Governments have implemented all sorts of tactics to try and mitigate climate change, most of which are expensive, require land use, and are not particularly powerful weapons against carbon. Therefore, seaweed aquaculture is ripe with untapped potential. Taking some of the strain of world-wide agriculture off the land benefits CO2 reduction, and provide an extra food source for humanity. Macroalgae has also been investigated as an alternative energy source to fossil fuels, and an alternative to livestock feed. Using macroalgae for these purposes would greatly reduce CO2 production from fuel burning and livestock. Seaweed farming, if supported is a fully sustainable and carbon neutral industry with the ability to be established in a variety of environments across the world.

When first studying the replication of DNA, it was unsure in way the DNA was actually replicated. The three models that were studied by scientitsts were the conservative, semiconservative, and dispersive model for DNA replication. The conservative model shows that two newly synthesized molecules of DNA come together while the original helix is joined back together. The semiconservative model shows that each parental strand is a template strand for the newly synthesized DNA molecule and one new strand is joined to one parental strand. The dispersive model shows that parental strands are dispersed into two new double helices. After experiements done by scientists, we know today that the DNA replication is semiconservative.

Yes, I think the title describes the subject of the paper well. The abstract summarizes the essential parts of the study and gave a brief description of the paper in a nutshell. It was also at the appropriate length. The plot sizes need to be explained more thoroughly, as this section has a large amount of given proportions without much information about the locations. There are no inaccuracies in this area. I do not think anything needs to be omitted, just a more thorough explanation of the sites. Each table and figure clearly presented important results. The tables are crammed together and need a larger space between them. This would make the tables and figures more effective. Data is not given in more than one place and units, standard errors, deviations, axis labels and legends are present. The legends are in the right format but are not on their own page. All the figures and tables seem necessary for the paper and no other figures or tables need to be added.