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| <b>Members:</b> Celine Siu Ham Zhang, Jiangshan Gao, Sizhe Duan, Marco Xue, Yeqi Wong, Xinyuan Wang, Bert Shan, James Haorui Wang, Junfei (Sophie) Li, Yiming Tang, Shao Chang, Siyuan Wang, Bohan Zhang<br> | | <b>Members:</b> Celine Siu Ham Zhang, Jiangshan Gao, Sizhe Duan, Marco Xue, Yeqi Wong, Xinyuan Wang, Bert Shan, James Haorui Wang, Junfei (Sophie) Li, Yiming Tang, Shao Chang, Siyuan Wang, Bohan Zhang<br> |
| <b>Primary PI:</b> Shiqi Wang.<br> | | <b>Primary PI:</b> Shiqi Wang.<br> |
− | <b>Instructors:</b> Xing (Leaves) Zhang, Tianze Zhu)<br> | + | <b>Instructors:</b> Xing (Leaves) Zhang, Tianze Zhu<br> |
| <b>Advisors:</b> Zhimeng Xu, Jun Ma, Aiqi Zhao, Li TianHong, Luo Shaowei, Weizhao Chen<br><br> | | <b>Advisors:</b> Zhimeng Xu, Jun Ma, Aiqi Zhao, Li TianHong, Luo Shaowei, Weizhao Chen<br><br> |
| All of us welcome you to visit and critique our poster!<br><br> | | All of us welcome you to visit and critique our poster!<br><br> |
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| <div class="title">Introduction & Inspiration</div> | | <div class="title">Introduction & Inspiration</div> |
| <div class="text"> | | <div class="text"> |
− | Our project is a continuation of the project created by QHFZ-China | + | Our project is a continuation of the project created by QHFZ-2019. Last year, we made a portable household uric acid detector, which is a genetically engineered bacteria. It would facilitate the recovery of gout patients. However, we realized that there are still some problems hindering its practical application. Among them, we wanted to solve the issue of room-temperature storage before anything else.<br> |
− | 2019. Last year, we made a portable household uric acid detector, which is a genetically
| + | |
− | engineered bacteria. It would facilitate the recovery of gout patients. However, we realized
| + | |
− | that there are still some problems hindering its practical application. Among them, we
| + | |
− | wanted to solve the issue of room-temperature storage before anything else. <br>
| + | |
| <img src="https://static.igem.org/mediawiki/2020/2/27/T--QHFZ-China--Poster_section1_1.png" | | <img src="https://static.igem.org/mediawiki/2020/2/27/T--QHFZ-China--Poster_section1_1.png" |
| alt=""><br> | | alt=""><br> |
− | Engineered bacteria need to be able to store for an extended period | + | Engineered bacteria need to be able to store for an extended period before being used at room temperature. They should stay alive while their metabolism paused. In laboratories, we use ultralow temperature to achieve this. That is why -80℃ refrigerator is necessary for molecular biology labs. However, it is hard to utilize such professional equipment from laboratories since they cannot be found daily. In general, household refrigerators can only achieve about -20℃, which is insufficient. Besides, it is not ideal to use the refrigerators to store bacteria, which we primarily use to store food. However, many engineered bacteria are designed for our daily life. We interviewed ten iGEM teams, and most of them believed that a suitable method to solve the problem would benefit the practical application of their products.<br> |
− | before being used at room temperature. They should stay alive while their metabolism paused.
| + | |
− | In laboratories, we use ultralow temperature to achieve this. That is why -80℃ refrigerator
| + | |
− | is necessary for molecular biology labs. However, it is hard to utilize such professional
| + | |
− | equipment from laboratories since they cannot be found daily. In general, household
| + | |
− | refrigerators can only achieve about -20℃, which is insufficient. Besides, it is not ideal
| + | |
− | to use the refrigerators to store bacteria, which we primarily use to store food. However,
| + | |
− | many engineered bacteria are designed for our daily life. We interviewed 10 iGEM teams, and
| + | |
− | most of them believed that a suitable method to solve the problem would benefit the
| + | |
− | practical application of their products.<br>
| + | |
| <img src="https://static.igem.org/mediawiki/2020/2/2b/T--QHFZ-China--Poster_section1_2.png" | | <img src="https://static.igem.org/mediawiki/2020/2/2b/T--QHFZ-China--Poster_section1_2.png" |
| alt=""><br> | | alt=""><br> |
− | As a result, we decide to come up with a biopreservation method, | + | As a result, we decided to develop a biopreservation method, which could help store the engineered bacteria without professional equipment. |
− | which could help the storage of engineered bacteria get rid of professional equipment.
| + | |
| </div> | | </div> |
| </div> | | </div> |
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| <img src="https://static.igem.org/mediawiki/2020/4/42/T--QHFZ-China--Poster_2.png"> | | <img src="https://static.igem.org/mediawiki/2020/4/42/T--QHFZ-China--Poster_2.png"> |
| <div class="info"> | | <div class="info"> |
− | <div class="title">Tardigrade</div> | + | <div class="title">The Limitations of Current Methods and Our Solution</div> |
| <div class="text"> | | <div class="text"> |
− | <b>Current Storage Methods</b><br><br> | + | <b>Common Storage Methods</b><br><br> |
− | There are several bio-preservation methods for bacteria. | + | There are several bio-preservation methods for bacteria. Nevertheless, at research institutes, ultra-low temperature freezing is the most commonly used method for long-term storage. However, the storage relies on specialized equipments, including the ultralow temperature refrigerators (-70℃~-80℃). Ultralow temperature refrigerators are expensive, noisy, and consumes lots of electricity, so there is usually no such equipment outside of laboratories.<br> |
− | Nevertheless, at research institutes, ultra-low temperature freezing is the most commonly
| + | |
− | used method for long-term storage. The method is effective and reliable. However, the
| + | |
− | storage relies on specialized equipments, including the ultralow temperature refrigerators,
| + | |
− | because the storage temperature needs to be lower than -70℃~-80℃. Ultralow temperature
| + | |
− | refrigerators are expensive, noisy, and consumes lots of electricity, so there is usually no
| + | |
− | such equipment outside of a laboratory. <br>
| + | |
| <img src="https://static.igem.org/mediawiki/2020/d/d7/T--QHFZ-China--Poster-sec2_3.png" | | <img src="https://static.igem.org/mediawiki/2020/d/d7/T--QHFZ-China--Poster-sec2_3.png" |
| alt=""><br> | | alt=""><br> |
− | During another possible preservation technique, sand preservation, | + | Though there are some biopreservation means besides ultralow temperature, each method has shortcomings. This promoted us to initiate our project.<br><br> |
− | the bacteria can be very easily contaminated due to sand usage. Therefore, very detailed
| + | |
− | procedures need to be carried out by professionals with scientific training, which is
| + | |
− | impossible for the general public to use. Hence, it is crucial to find a method of bacterial
| + | |
− | storage suitable popularization among normal people. <br><br>
| + | |
| | | |
| <b>Freeze-drying / Lyophilization</b><br><br> | | <b>Freeze-drying / Lyophilization</b><br><br> |
− | However, compared to -80℃ storage, we found new methods that are | + | The metabolism slows down if the cells lose water. Indeed, there are some dried bacteria products on the market. Thus, we decided to make engineered bacteria into dry powder. Then the powder can be stored at room temperature without any professional equipment.<br> |
− | more applicable. From our textbooks, we knew that the metabolism slows down if the cells
| + | |
− | lose water. Indeed, there are some dried bacteria products on the market. Thus, we decided
| + | |
− | to make engineered bacteria into dry powder. Then the powder can be stored at room
| + | |
− | temperature without any professional equipment. <br>
| + | |
| <img src="https://static.igem.org/mediawiki/2020/f/f4/T--QHFZ-China--Poster-sec2_4.png" | | <img src="https://static.igem.org/mediawiki/2020/f/f4/T--QHFZ-China--Poster-sec2_4.png" |
| alt=""><br> | | alt=""><br> |
− | The direct drying method is time-consuming, and the metabolism will | + | The direct drying method is time-consuming, and the metabolism will not stop during the process. However, freeze-drying is very fast. Indeed, freeze-drying has been widely used to produce vaccines, medicaments, and food because it can maintain the materials' original state.<br> |
− | not stop during the process. However, freeze-drying is very fast. Firstly, the temperature
| + | |
− | decreases, and the metabolism slows down. After freezing, the water is quickly lost in the
| + | |
− | vacuum via sublimation. Indeed, freeze-drying has been widely used to produce vaccines,
| + | |
− | medicaments, and food because it can maintain the materials' original state. The dried form
| + | |
− | of the products, called "cakes," is porous and easy to be rehydrated. <br>
| + | |
| <img src="https://static.igem.org/mediawiki/2020/b/b5/T--QHFZ-China--Poster-sec2_5.png" | | <img src="https://static.igem.org/mediawiki/2020/b/b5/T--QHFZ-China--Poster-sec2_5.png" |
| alt=""><br> | | alt=""><br> |
− | Nonetheless, during the process, at least three stresses affect | + | Nonetheless, during the process, at least three stresses affect cells' survival: freeze, desiccation, and vacuum. Several studies have tried to improve the survival rate of bacteria. Firstly, researchers tried to improve the procedure of freeze-drying; Secondly, they tried to optimize the constituent of protectants and excipients in the solution before lyophilization, but the survival rate still stayed at about 10%. Thus, we hope to introduce another element to elaborate on previous studies.<br><br> |
− | cells' survival: freeze, desiccation, and vacuum. Several studies have tried to improve the
| + | |
− | survival rate of bacteria. Firstly, researchers tried to improve the procedure of
| + | |
− | freeze-drying; Secondly, they tried to optimize the constituent of protectants and
| + | |
− | excipients in the solution before lyophilization, but the survival rate still stayed at
| + | |
− | about 10%. Thus, we hope to introduce another element to elaborate on previous
| + | |
− | studies.<br><br>
| + | |
| | | |
| <b>Tardigrade (water bear) / TDPs</b><br><br> | | <b>Tardigrade (water bear) / TDPs</b><br><br> |
− | Therefore, to apply lyophilization/freeze-drying, we need to | + | To apply lyophilization/freeze-drying, we need to utilize the proteins of a very special organism. Tardigrade (water bear) is a miraculous organism found in 1773 and is famous for its strong vitality. It can survive extreme environments, like freezing, desiccation, vacuuming, extreme temperatures, high pressure, and radiation.<br> |
− | utilize the proteins of a very special organism. Tardigrade (water bear) is a miraculous
| + | |
− | organism found in 1773 and is famous for its strong vitality. It can survive extreme
| + | |
− | environments, like freezing, desiccation, vacuuming, extreme temperatures, high pressure,
| + | |
− | and radiation.<br>
| + | |
| <img src="https://static.igem.org/mediawiki/2020/c/c6/T--QHFZ-China--Poster-sec2_6.png" | | <img src="https://static.igem.org/mediawiki/2020/c/c6/T--QHFZ-China--Poster-sec2_6.png" |
| alt=""><br> | | alt=""><br> |
− | The strong vitality of tardigrades at least partially owes to TDPs | + | The strong vitality of tardigrades at least partially owes to TDPs (Tardigrade intrinsically Disordered Proteins), which are exceptionally resistant to desiccation.<br> |
− | (Tardigrade intrinsically Disordered Proteins), which are exceptionally resistant to
| + | |
− | desiccation.<br>
| + | |
| <img src="https://static.igem.org/mediawiki/2020/d/dc/T--QHFZ-China--Poster-sec2_7.png" | | <img src="https://static.igem.org/mediawiki/2020/d/dc/T--QHFZ-China--Poster-sec2_7.png" |
| alt=""><br> | | alt=""><br> |
− | TDPs are found in 2017, mainly including several Cytosolic-abundant | + | TDPs are found in 2017<sup>[1]</sup>, mainly including several Cytosolic-abundant heat soluble proteins (CAHS) and secreted massive heat soluble proteins (SAHS). TDPs were proven to protect living cells from drying and maintain the activeness of proteins after freeze-drying, but no one studied whether they can protect living cells from freeze-drying or not. Therefore, we tried to allow bacteria to express TDPs and tested whether they can survive freeze-drying and the subsequent storage processes.<br> |
− | heat soluble proteins (CAHS) and secreted massive heat soluble proteins (SAHS). TDPs were
| + | |
− | proven to protect living cells from drying and maintain the activeness of proteins after
| + | |
− | freeze-drying, but no one studied whether they can protect living cells from freeze-drying
| + | |
− | or not. Therefore, we tried to allow bacteria to express TDPs and tested whether they can
| + | |
− | survive freeze-drying and the subsequent storage processes. <br>
| + | |
| <img src="https://static.igem.org/mediawiki/2020/2/21/T--QHFZ-China--Poster-sec2_8.png" | | <img src="https://static.igem.org/mediawiki/2020/2/21/T--QHFZ-China--Poster-sec2_8.png" |
| alt=""><br> | | alt=""><br> |
− | In this project, we studied 5 TDPs: CAHS 89226, CAHS 94205, CAHS | + | In this project, we studied 5 TDPs: CAHS 89226, CAHS 94205, CAHS 106094, CAHS 107838, and SAHS 33020, as well as two gene parts that are used before TDPs were found: LEA (Late Embryogenesis Abundant protein) from <i> Glycine max </i><sup>[5]</sup> and OtsBA (producing trehalose)<sup>[4]</sup>.<br><br> |
− | 106094, CAHS 107838, and SAHS 33020, as well as two gene parts that are used before TDPs
| + | |
− | were found: LEA (Late Embryogenesis Abundant protein) from <i> Glycine max </i> and OtsBA
| + | |
− | (producing trehalose) <br><br>
| + | |
− | <div class="subsection two_thirds" id="sb1">
| + | |
− | <img src="https://static.igem.org/mediawiki/2020/e/ea/T--QHFZ-China--Poster_section2.png">
| + | |
− | </div>
| + | |
− | <div class="subsection third" id="sb2">
| + | |
− | fksajhfkasdklasjdlkasjdlaksjdlkasjdlksajdlkajd;asjd;lasjdlasjd<br>
| + | |
− | <br>
| + | |
− | sss
| + | |
− | </div>
| + | |
− | <div class="clear"></div>
| + | |
− | dhfkldsjflkasjf;ljdasl;kfd;laskdf;lsakf;ljslfjdlas;jf;asjdfl
| + | |
| </div> | | </div> |
| </div> | | </div> |
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| <b>Members:</b> Celine Siu Ham Zhang, Jiangshan Gao, Sizhe Duan, Marco Xue, Yeqi Wong, Xinyuan Wang, Bert Shan, James Haorui Wang, Junfei (Sophie) Li, Yiming Tang, Shao Chang, Siyuan Wang, Bohan Zhang<br> | | <b>Members:</b> Celine Siu Ham Zhang, Jiangshan Gao, Sizhe Duan, Marco Xue, Yeqi Wong, Xinyuan Wang, Bert Shan, James Haorui Wang, Junfei (Sophie) Li, Yiming Tang, Shao Chang, Siyuan Wang, Bohan Zhang<br> |
| <b>Primary PI:</b> Shiqi Wang.<br> | | <b>Primary PI:</b> Shiqi Wang.<br> |
− | <b>Instructors:</b> Xing (Leaves) Zhang, Tianze Zhu)<br> | + | <b>Instructors:</b> Xing (Leaves) Zhang, Tianze Zhu<br> |
| <b>Advisors:</b> Zhimeng Xu, Jun Ma, Aiqi Zhao, Li TianHong, Luo Shaowei, Weizhao Chen<br><br> | | <b>Advisors:</b> Zhimeng Xu, Jun Ma, Aiqi Zhao, Li TianHong, Luo Shaowei, Weizhao Chen<br><br> |
| All of us welcome you to visit and critique our poster!<br><br> | | All of us welcome you to visit and critique our poster!<br><br> |