The paper proposes an experimental research to examine if the molecular beacons can be usefully employed in the detection of the gene expression of the cancer cells. In this regard there will be an experimental research conducted on the cancer diagnosed stomach and a normal stomach by injecting the molecular beacons on both of the stomachs and then checking the reactions in both of them. The proposed research will be a continuation of the experimental researchers done so far to detect the gene expression in the cancer cell by using the molecular beacon. It is found from the early researches that the injection of the molecular beacons in the cancer affected cell produces strong fluorescent as compared with the normal cell. Thus the different reaction of the molecular beacon injecting in the cancer affected cells show that the molecular beacon injection generates different type of reaction in the cancer cell so it can help in the detection of the gene expression in the caner cell. Thus the molecular beacons can be used for the testing of the cells to check the indications of cancer and further in the treatment of the disease.
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Statement of Problem
Molecular Beacons have been used in the fields of biology and chemistry for performing a wide range of functions and experiments. Molecular Beacons are the fluorescent and oligonucleotide based probes that have exceptional sensitivity as well as specificity (Tyagi et al, p303). Due to this reasons the researchers and the biology and chemistry experts concentrate to perform different experiments through which the attributed of the molecular beacons can be successfully used in the detection of the gene expression. Many of the researches came up with the conclusions that the molecular beacons have the capability to react in a different manner in the cancer cells as compared with the normal cell. This was an important development and since then the researchers focus to find out that how they can use the molecular beacons for the detection of the gene expression in the cancer cell. In this regard there are some researchers already conducted (e.g. Xiang et al, p2) that proves the different type of reactions of the molecular beacons in the cells. Thus in continuation with the existing researches, the proposed research will conduct an experiment on the animal cells to check the reaction of injecting the molecular beacon in the normal and cancer cells. The main focus of the proposed experimental research will be to find out the question of the following research question:
“How molecular beacons can be used for the detection of gene expression in the cancer cells and how the molecular beacons can help in the cure of the cancer in living cells”
Need and Significance of the Research
There is great significance of this research because the possible outcome of the research will pave new ways for the biologist to find out the ways of detecting and curing the caner in the living cells. Many of the cases of the cancer are not diagnosed properly at the early stages due to which the patient loose the possibility of complete cure and treatment of the disease but if the molecular bacons can be determined as the useful way of detection the gene expression in the cancer cells then it will become easy to diagnose the cancer in the early stages and then it can further help in the treatment of the disease as well.
The proposed research will be conducted in two phases. In the first phase the review of the related literature will be conducted to have deep understanding of the related terms, issues and concepts. In this regard the existing researches done around the same topic will also be studied to find appropriate way of conducting the experiment and in the light of the review of the related researchers and literature the second phase of the research i.e. the practical will be conducted. In the second and the experimental phase two living cells of the stomach of an animal will be taken and the specific ratio of the molecular beacons will be injected in both of the cells. One of the cells will be normal whereas the other will be a cancer diagnosed cell. Both of these cells will be monitored for specific time period and the changes occurred in both of the cells during that time period will be measured to find out that how the molecular beacons differently reacts in the normal and cancer cells. The molecular beacons will not only be injected to detect the gene expression but also to find out that how these can help in the treatment of the cancer cell.
Brief Review of the Literature
Dirk (2001) explains that the development of the molecular beacons in the year 1996 provided the biologists and the chemistry experts several new ways of using these molecular beacons in their research work for wide range of purposes (p11). Since then these have been used for different types of experiments especially for the detection of the gene expressions. It is found that the molecular beacons can play a significant role in the detection of the cancer cell genomics and the reactions of the molecular beacons after entering the cancer cell can help in the detection of the disease significantly (Noya, p299)
Though the above literature in very brief but the dissertation will provide detailed account of information about all the topics, researches and concepts related with the topic.
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Structure of the Dissertation
The proposed dissertation will consist of following parts
- Introduction: The first chapter will introduce the main concept of the research, the motive behind conducting the experimental study, its need, significance and possible contribution to the literature and related field.
- Literature Review: All the concepts, issues, theories and models associated with the topic of the research will be explained in this chapter with the help of the review of the existing research studies conducted around the topic
- Methodology: This chapter will explain in details the ways of conducting the experimental research. In the regard all the minor and major details of the methodology will be described in this chapter
- Experiment Results: This chapter will contain all the information about the outcome of the experimental study. It will show the results obtained from the monitoring of both the cells during the specific time period.
- Analysis and Discussion: In this chapter the results or outcome of the experimental research will be analyzed and then discussed with the support of the concepts and theories studies in the review of the literature
- Conclusion: This chapter will summarize the entire research study and will provide the detailed information of the objectives and outcome of the study to conclude the entire work.
Proposed Reference List
A. K. Chen, M. A. Behlke, and A. Tsourkas (2007), Avoiding false-positive signals with nuclease-vulnerable molecular beacons in single living cells, Nucleic Acids Res. Web.
Bratu,D.P., Cha,B.J., Mhlanga,M.M., Kramer,F.R. and Tyagi,S. Visualizing the distribution and transport of mRNAs in living cells. Proc.Natl.Acad.Sci.U.S.A. (2003), 100, 13308-13313.
Cui,Z.Q., Zhang,Z.P., Zhang,X.E., Wen,J.K., Zhou,Y.F. and Xie,W.H. Visualizing the dynamic behavior of poliovirus plus-strand RNA in living host cells. Nucleic Acids Res. (2005), 33, 3245-3252.
Dirks, Roeland W.; Molenaar, Chris; Tanke, Hans J. (2001), Methods for visualizing RNA processing and transport pathways in living cells., Histochemistry and Cell Biology 115(1), 3-11.
Golzio,M., Rols,M.P. and Teissie,J. (2004), In vitro and in vivo electric field-mediated permeabilization, gene transfer, and expression. Methods 33, 126-135.
Nitin, Nitin; Santangelo, Philip J.; Kim, Gloria; Nie, Shuming; Bao, Gang. Peptide-linked molecular beacons for efficient delivery and rapid mRNA detection in living cells, Nucleic Acids Research (2004), 32(6), e58/1-e58/8.
Noya O; Patarroyo M E; Guzman F; Alarcon de Noya B (2003), Immunodiagnostic of parasitic diseases with synthetic peptides. Current protein & peptide science 4(4), pp299-308.
Perlette,J. and Tan,W.H., Real-time monitoring of intracellular mRNA hybridization inside single living cells. Anal.Chem. (2001), 73, 5544-5550.
Piatek, Amy S.; Tyagi, Sanjay; Pol, Arno C.; Telenti, Amalio; Miller, Lincoln P.; Kramer, Fred Russell; Alland, David (1998), Molecular beacon sequence analysis for detecting drug resistance in Mycobacterium tuberculosis. Nature Biotechnology 16(4), pp359-363.
Santangelo, Philip J.; Nix, Brent; Tsourkas, Andrew; Bao, Gang. Dual FRET molecular beacons for mRNA detection in living cells. Nucleic Acids Research (2004), 32(6), e57/1-e57/9.
Tyagi, Sanjay; Kramer, Fred Russell (1996), Molecular beacons: probes that fluoresce upon hybridization, Nature Biotechnology 14(3), pp303-8.
Tan, Weihong; Wang, Kemim; Drake, Timothy J (2004), Molecular beacons, Current Opinion in Chemical Biology 8(5), pp547-553.
Xiang-Hong Peng, Ze-Hong Cao, Jin-Tang Xia, Grant W. Carlson, Melinda M. Lewis, William C. Wood and Lily Yang (2005), Real-time Detection of Gene Expression in Cancer Cells Using Molecular Beacon Imaging: New Strategies for Cancer Research, Cancer Research 65, 1909-1917, March 1, 2005, American Association for Cancer Research.