Christine
Curtis presents this Friday during seminar. The title of her talk is:
"Recombineering
strategies for isolating, cloning, and deleting genes in Enterobacter Species
YSU and S. maltophilia OR02"
She has
provided links to the following PDF documents as background reading:
Bring your lunch and enjoy yet another excellent
seminar, I'm sure (No pressure, Christine!!!!).
First of all, sorry for being so late with this. But I hope everyone had a good break!
ReplyDeleteChristine’s work includes the use of the lambda-red bacteriophage to perform recombineering in different bacteria strains such as Enterobacter YSU and S. maltophilia. Recombineering can be an alternative to restriction digestion and ligation for gene alteration. Christine developed a custom plasmid, transformed it in bacteria cultures, and amplified certain PCR products. Lastly she is using the lambda-red phage to interrupt a mercury resistance gene in the Enterobacter YSU bacteria.
What possible applications could recombineering, or other somewhat similar methods such as CRISPR/Cas9, be utilized for in gene therapy? In particular, for the prevention or treatment of disease.
I just read an article about how they are using gene therapy to help individuals that are diagnosed with hemophilia. Hemophilia is often caused by the mutation within the factor IX (FIX) gene. In 2009, a Doctor and his colleagues came across an Italian man that had a condition known as thrombosis. This condition is characterized by excessive blood clotting within blood vessels. Researchers discovered that this patient had a genetic mutation within the FIX gene that promoted thrombosis. It was the first and only FIX gene mutant that has been found not to cause hemophilia. Upon isolating this gene, vectors were used to introduce this mutant gene into canines that exhibited symptoms of hemophilia. They found that it was effective in treating the severity of their hemophilia (taking it from sever to mild). Similar results were found within studies done on mice.
ReplyDeleteGene therapy is still a highly controversial idea, because one can pass along an "artificially acquired mutation". However there are diseases caused not simply by just a mutation but how that mutation is expressed. For example, Fragile X Tremor Ataxia syndrome (FXTAS) is more severe upon the increased expression of the mutated FMR1 gene which produces a protein that plays a role in regulation of other proteins. Recombineering can used to increase regulatory elements which may cascade onto the transcription of the defective gene.
ReplyDeleteGenetic diseases like cystic fibrosis, which is caused by mutations to a type of cell surface chloride channel, can be knocked out with the help of recombineering techniques similar to those Christine talked about.
ReplyDeleteA research team lead by Dr. Hans Sollinger at the University of Wisconsin School of Medicine and Public Health are developing a gene therapy for Type 1 diabetes mellitus (T1DM). The objective of this research is to find a good substitute that would effectively manage Diabetes mellitus and as well it would eliminate the need for daily insulin shots and provide a way to better regulate glucose metabolism.
ReplyDeleteChristine’s seminar presentation was an interesting look into both the history of and the current iterations of recombineering techniques in bacterial organisms. Her results show the potential for efficient and mass production of unique, modified bacteria that could help explore genetic function and the understanding of different biological pathways.
ReplyDeleteA YSU alumnus in currently at the University of Utah School of Medicine has helped to pioneer a medical technique that delivers a gene, SDF-1, to the heart after an individual has suffered a heart attack to help promote healing. While not specifically recombineering, the technique does take adavantage of controlling genes to help promote advanced healing.
When looking into this type of research and how it can be used in humans I came upon a study where the researchers were looking at ways to treat Huntingtons disease. This neurological degenerative disease which is caused by the expansion of the CAG repeats in the huntintin protein. The more repeats present the earlier the onset of the disease. Human induced pluripotent stem cells (iPSCs) derived from HD patient fibroblasts were targeted for correction by the replacement of the expanded CAG repeat with a normal repeat using homologous recombination. The corrected iPSCs were able to successfully differentiate into neurons and the HD pathogenic signaling pathways were successfully normalized, reversing the disease phenotypes such as susceptibility to cell death and altered mitochondrial bioenergetics in neural stem cells.
ReplyDeletehttp://www.sciencedirect.com/science/article/pii/S1934590912003372
Christine’s presentation was a great look into the development and refining of molecular genetic techniques. Her work would be highly useful both in research labs and teaching labs to show students how mutations can affect bacterial organisms.
ReplyDeleteRecombineering techniques presented by Muck-Hausl et al. in 2015 use recombinant adenoviruses containing a double-stranded DNA genome that have potential uses for gene therapy. They can modify these recombinant adenoviral vectors (AdVs) genetically for use not only specific gene therapies but also future vaccination studies.
Christine's presentation showed the positive effects of recombineering as well as the several difficulties in developing this technique. Once this recombineering is perfected, it will save a lot of time and money in the microbiology labs.
ReplyDeleteAccording to the American Society of Gene & Cell Therapy, 2/3 of all gene therapy is designed to treat cancer cells. Specifically there is one methlled suicide therapy of engineering called suicide therapy, which is used to insert a kill gene into the specific cancer cells. This same technique is also being developed to treat HIV/AIDS and Hepatitis viruses.
Good job Christine on your presentation.
ReplyDeleteAutoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a rare autoimmune disease that primarily affects the endocrine glands caused by a mutation in the autoimmune regulator (AIRE) gene. Researchers are exploring technical aspects and limitations of the procedure behind recombineering in gene therapy.
http://www.bloodjournal.org/content/99/8/2694?sso-checked=true
http://scitechnol.com/gene-therapy-in-primary-immunodeficiencies-Ub9V.php?article_id=1281
Nicely done folks!!!!!
ReplyDelete