Difference between revisions of "Team:Shanghai HS ID/Description"

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<h3>★  ALERT! </h3>
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<p>This page is used by the judges to evaluate your team for the <a href="https://2021.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2021.igem.org/Judging/Awards"> award listed below</a>. </p>
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2021.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
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<h1>Project Description </h1>
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<h3>Bronze Medal Criterion #3</h3>
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<p>Describe how and why you chose your iGEM project.
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Please see the <a href="https://2021.igem.org/Judging/Medals">2021 Medals Page</a> for more information.
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<h3>What should this page contain?</h3>
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<ul>
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<li> A clear and concise description of your project.</li>
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<li>A detailed explanation of why your team chose to work on this particular project.</li>
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<li>References and sources to document your research.</li>
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<li>Use illustrations and other visual resources to explain your project.</li>
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<h3>Inspiration</h3>
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<p>See how other teams have described and presented their projects: </p>
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<ul>
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<li><a href="https://2019.igem.org/Team:Leiden/Description">2019 Leiden</a></li>
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<li><a href="https://2019.igem.org/Team:ITESO_Guadalajara/Description">2019 ITESO Guadalajara</a></li>
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<li><a href="https://2020.igem.org/Team:Technion-Israel/Description">2020 Technion Israel</a></li>
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<li><a href="https://2020.igem.org/Team:Botchan_Lab_Tokyo/Description">2020 Botchan Lab Tokyo</a></li>
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<li><a href="https://2020.igem.org/Team:St_Andrews/Description">2020 St Andrews</a></li>
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<li><a href="https://2020.igem.org/Team:MIT/Description">2020 MIT</a></li>
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<h3>Advice on writing your Project Description</h3>
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We encourage you to put up a lot of information and content on your wiki, but we also encourage you to include summaries as much as possible. If you think of the sections in your project description as the sections in a publication, you should try to be concise, accurate, and unambiguous in your achievements. Your Project Description should include more information than your project abstract.
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<h3>References</h3>
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<p>iGEM teams are encouraged to record references you use during the course of your research. They should be posted somewhere on your wiki so that judges and other visitors can see how you thought about your project and what works inspired you.</p>
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        <div class="article-title">Lactobacillus Casei</div>
 +
        <div class="article-content">Lactobacillus caseiis a lactic acid bacterium that could be found in various
 +
            natural or man-made environments <b>[1]</b>. It is used to make numerous day-to-day products including
 +
            cheese,
 +
            yogurt, and other dairy beverages. A study conducted in 2010 show that probiotics like L.Caseido contain
 +
            health promoting properties. Thirty-four adults were divided into two groups – one with a four-week intake
 +
            of probiotic fermented beverage, and one without. The results suggest that "probiotic fermented milk
 +
            beverage has an intestine-conditioning effect by improving the frequency of defecation and stool quality and
 +
            increasing the intrinsic bifidobacteria in healthy individuals with soft stool" <b>[7]</b>. As a probiotic,
 +
            L.caseiis also much safer and beneficial to the human body as a carrier for genetic materials than the
 +
            widely used E.colibacteria. <b>[3]</b>Making L.Caseihighly applicable in many frontiers of medicine,
 +
            pharmaceuticals, and food. However due to their restriction-modification system the bacteria’s
 +
            transformation rate is very inefficient, hence eminently restraining their capabilities.</div>
 +
        <div class="article-title">Obstacles</div>
 +
        <div class="article-content">L.Caseihas a self-protection mechanism called the restriction-modification system.
 +
            This system protects the bacteria through using restriction endonucleases to disable the invasive external
 +
            DNAs. However this leads to a low transformation efficiency when trying to introduce external DNAs in order
 +
            to maximizeL.Casei'scapabilities <b>[2]</b> This inefficiency hinders the study of other applications of
 +
            L.caseiin
 +
            fields such as probiotic-delivered vaccines<b>[3]</b>and other medicines. We discovered that traditional
 +
            methods
 +
            such as methylating foreign DNA or using high-temperatures to temporarily denature the restriction
 +
            modification system are not only time-consuming and costly but also not solving the problem fundamentally
 +
            <b>[4]</b><b>[3]</b>. Inspired by genetic engineering tools, our team came up with a new way of utilizing
 +
            CRISPR-Cas9 to
 +
            thoroughly solve this problem.
 +
        </div>
 +
        <div class="article-title">Solution</div>
 +
        <div class="article-content">We constructed a plasmid equipped with a CRISPR-Cas9 complex that cuts and removes
 +
            a selected segment of the L. casei DNA. Our target gene is LSEI-2094. This gene is involved in the synthesis
 +
            of an enzyme that is essential in the restriction-modification system [5]. The cell's efforts to repair its
 +
            DNA induces mutations, which help to inactivate the gene and prevent the production of the enzyme. After
 +
            this modification foreign DNAs could enter and express themselves much more efficiently in L.Casei.With a
 +
            strain of bacteria ready to be tailored to our needs, the benefits are immeasurable. The benefits of L.
 +
            caseicould come to fruition if the technology is applied to popular and impactful industries such as
 +
            pharmaceuticals and food. </div>
 +
        <div class="article-title">
 +
            Reference
 +
        </div>
 +
        <div class="article-content">1. Buriti, F. C., & Saad, S. M. (2007). Bactérias do grupo Lactobacillus casei:
 +
            caracterização, viabilidade como probióticos em alimentos e sua importância para a saúde humana [Bacteria of
 +
            Lactobacillus casei group: characterization, viability as probiotic in food products and their importance
 +
            for human health]. Archivos latinoamericanos de nutricion, 57(4), 373–380.</div>
 +
        <div class="article-content">2. Matsumoto, K., Takada, T., Shimizu, K., Moriyama, K., Kawakami, K., Hirano, K.,
 +
            Kajimoto, O., & Nomoto, K. (2010). Effects of a probiotic fermented milk beverage containing Lactobacillus
 +
            casei strain Shirota on defecation frequency, intestinal microbiota, and the intestinal environment of
 +
            healthy individuals with soft stools. Journal of bioscience and bioengineering, 110(5), 547–552.
 +
            https://doi.org/10.1016/j.jbiosc.2010.05.016</div>
 +
        <div class="article-content">3. National Center for Biotechnology Information (NCBI)[Internet]. Bethesda (MD):
 +
            National Library of Medicine (US), National Center for Biotechnology Information;1988. Available from:
 +
            https://www.ncbi.nlm.nih.gov/</div>
 +
        <div class="article-content">4. Walsh, C. J., Healy, S., O'Toole, P. W., Murphy, E. F., & Cotter, P. D. (2020).
 +
            The probiotic L. casei LC-XCAL™ improves metabolic health in a diet-induced obesity mouse model without
 +
            altering the microbiome. Gut microbes, 12(1), 1704141. https://doi.org/10.1080/19490976.2020.1747330</div>
 +
        <div class="article-content">5. 汪川 & 张朝武.(2008).以益生菌为载体的基因工程疫苗研究进展. 卫生研究(01),118-122.
 +
            doi:CNKI:SUN:WSYJ.0.2008-01-045.</div>
 +
        <div class="article-content">6. 潘荣荣. 乳酸杆菌破壁方法及限制——修饰系统对转化效率影响的研究[D]. 扬州大学.
 +
            https://kns.cnki.net/KCMS/detail/detail.aspx?dbname=CMFD201801</div>
 +
        <div class="article-content">7. Desirazu, N, Rao, et al. Type III restriction-modification enzymes: a historical
 +
            perspective[J]. Nucleic Acids Research, 2013, 42(1):45. https://doi.org/10.1093/nar/gkt616</div>
 +
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Revision as of 15:14, 15 October 2021

Shanghai_HS_ID

Project Description
Lactobacillus Casei
Lactobacillus caseiis a lactic acid bacterium that could be found in various natural or man-made environments [1]. It is used to make numerous day-to-day products including cheese, yogurt, and other dairy beverages. A study conducted in 2010 show that probiotics like L.Caseido contain health promoting properties. Thirty-four adults were divided into two groups – one with a four-week intake of probiotic fermented beverage, and one without. The results suggest that "probiotic fermented milk beverage has an intestine-conditioning effect by improving the frequency of defecation and stool quality and increasing the intrinsic bifidobacteria in healthy individuals with soft stool" [7]. As a probiotic, L.caseiis also much safer and beneficial to the human body as a carrier for genetic materials than the widely used E.colibacteria. [3]Making L.Caseihighly applicable in many frontiers of medicine, pharmaceuticals, and food. However due to their restriction-modification system the bacteria’s transformation rate is very inefficient, hence eminently restraining their capabilities.
Obstacles
L.Caseihas a self-protection mechanism called the restriction-modification system. This system protects the bacteria through using restriction endonucleases to disable the invasive external DNAs. However this leads to a low transformation efficiency when trying to introduce external DNAs in order to maximizeL.Casei'scapabilities [2] This inefficiency hinders the study of other applications of L.caseiin fields such as probiotic-delivered vaccines[3]and other medicines. We discovered that traditional methods such as methylating foreign DNA or using high-temperatures to temporarily denature the restriction modification system are not only time-consuming and costly but also not solving the problem fundamentally [4][3]. Inspired by genetic engineering tools, our team came up with a new way of utilizing CRISPR-Cas9 to thoroughly solve this problem.
Solution
We constructed a plasmid equipped with a CRISPR-Cas9 complex that cuts and removes a selected segment of the L. casei DNA. Our target gene is LSEI-2094. This gene is involved in the synthesis of an enzyme that is essential in the restriction-modification system [5]. The cell's efforts to repair its DNA induces mutations, which help to inactivate the gene and prevent the production of the enzyme. After this modification foreign DNAs could enter and express themselves much more efficiently in L.Casei.With a strain of bacteria ready to be tailored to our needs, the benefits are immeasurable. The benefits of L. caseicould come to fruition if the technology is applied to popular and impactful industries such as pharmaceuticals and food.
Reference
1. Buriti, F. C., & Saad, S. M. (2007). Bactérias do grupo Lactobacillus casei: caracterização, viabilidade como probióticos em alimentos e sua importância para a saúde humana [Bacteria of Lactobacillus casei group: characterization, viability as probiotic in food products and their importance for human health]. Archivos latinoamericanos de nutricion, 57(4), 373–380.
2. Matsumoto, K., Takada, T., Shimizu, K., Moriyama, K., Kawakami, K., Hirano, K., Kajimoto, O., & Nomoto, K. (2010). Effects of a probiotic fermented milk beverage containing Lactobacillus casei strain Shirota on defecation frequency, intestinal microbiota, and the intestinal environment of healthy individuals with soft stools. Journal of bioscience and bioengineering, 110(5), 547–552. https://doi.org/10.1016/j.jbiosc.2010.05.016
3. National Center for Biotechnology Information (NCBI)[Internet]. Bethesda (MD): National Library of Medicine (US), National Center for Biotechnology Information;1988. Available from: https://www.ncbi.nlm.nih.gov/
4. Walsh, C. J., Healy, S., O'Toole, P. W., Murphy, E. F., & Cotter, P. D. (2020). The probiotic L. casei LC-XCAL™ improves metabolic health in a diet-induced obesity mouse model without altering the microbiome. Gut microbes, 12(1), 1704141. https://doi.org/10.1080/19490976.2020.1747330
5. 汪川 & 张朝武.(2008).以益生菌为载体的基因工程疫苗研究进展. 卫生研究(01),118-122. doi:CNKI:SUN:WSYJ.0.2008-01-045.
6. 潘荣荣. 乳酸杆菌破壁方法及限制——修饰系统对转化效率影响的研究[D]. 扬州大学. https://kns.cnki.net/KCMS/detail/detail.aspx?dbname=CMFD201801
7. Desirazu, N, Rao, et al. Type III restriction-modification enzymes: a historical perspective[J]. Nucleic Acids Research, 2013, 42(1):45. https://doi.org/10.1093/nar/gkt616
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