Difference between revisions of "Team:BGIC-Union"

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<h1>Hi, I am dCasentry,</h1>
 
<h1>Hi, I am dCasentry,</h1>
 
<h2>a T7—dCas9 multi-output DNA sensor designed to detect ctDNA!</h2>
 
<h2>a T7—dCas9 multi-output DNA sensor designed to detect ctDNA!</h2>
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<p>Cancer is a leading cause of death worldwide, accounting for 8.8 million deaths in 2015, and the most common causes of cancer death is lung cancer (1.69 million deaths).
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<p>Cancer is a leading cause of death worldwide, accounting for 8.8 million deaths in 2015, and the most common cause of cancer death is lung cancer (1.69 million deaths). Nowadays, multiple lung cancer detection methods are developed, one of them being liquid biopsy. The technique of liquid biopsy is much better than the popular biopsy as it obviates pain and complication incurred to patients by detecting various tumor markup, including short strand circulating tumor DNA (CtDNA) in the blood. However, it still requires laboratory apparatus such as PCR instrument.</p>
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Nowadays, multiple lung cancer detection methods are developed, and I can be an assistant in liquid biopsy whose detection minimizes  trauma and pain. The tumor marker I detect is circulating tumor DNA (ctDNA), tumor-derived fragmented DNA in the bloodstream that is not associated with cells. It is released by the dying cells during the biological process of apotheosis and necrosis, or active release from viable tumor cells.
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<p>Thus our primary task is to devise a more efficient, rapid and convenient lung cancer prognosis method. We construct a paper chip containing powerful biosensor capable of detecting the fusion gene, EML4-ALK, which only exists only in NSCLC (non small cell lung cancer) patients’ blood. Specific targeted drug for this mutation has been produced, which means patients can receive precise treatment as soon as possible when the detection result is positive. </p>
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<p>The biosensor, dCasentry, consists of paired dCas9 protein guided by sgRNA and linked with split T7 RNA polymerase that serves as a switch. When the targeting lung cancer mutation gene is detected, visible signal will be released as the T7 RNA polymerase starts transcription. DCasentry not only deals with low ctDNA concentration in blood but also capable of producing various kinds of signal output, such as GFP and RFP.
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<p>I am an improved version of the paired dCas9 system designed by 2015 Peking university iGEM, composing of sgRNA, a pair of dCas9 protein and a split T7 RNA polymerase switch connected to each dCas9 protein via a linker. The sgRNA is designed to detect target genes and I will complete my task according to this order: First, the sgRNA will instruct each sgRNA-dCas9 complex to certain locus. Next, unlike Peking University, a T7 RNA polymerase rather than a luciferase is split apart and connected to dCas9 protein via a linker. I call them NT7-dCas9 and CT7-dCas9. Once two sgRNA are attached to the target sequence, two split parts will reunion and reassemble to form a complete RNA polymerase and start transcription consistently, which means even though the initial ctDNA concentration is low, the signal will continue to be magnified until it can be observed. Also, there is no limit on the content I can transcribed, that is, I am able to give out many kinds of signals, including GFP, RFP or LacZ. </p>
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<p>In this project my main goal is to detect the fusion gene <i>EML4-ALK</i>. Some of the cancer, including non small cell lung cancer (NSCLC), is caused by gene fusion. When two respective original genes fuse together, it produces the expression product of fusion gene, which induced the canceration of cells. EML4-ALK exists only in NSCLC patients’ blood which prevents false positive result during detection and there is a specific  targeted drug for this mutation which means patients can have precise treatment as soon as possible when the detection result is positive. My work is meaningful.</p>
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<p> We adopt freeze-dried paper chip as the vector of the detection system to simplify its operation as well as storage. The DNA device and proteins needed for detection are mixed with cell-free system and freeze-dried on the paper chip. During testing, the blood sample of the patient provides a liquid environment to reactivate the cell-free system. The product will be presented as a kit, containing including hemostix, a hand power centrifuge, NASBA reaction system and the paper screening chip, that can be widely used on the medical field. </p>
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<p>Finally, as a well-rounded and efficient lung cancer fusion gene detection sentry, I am proud to point out this important reason why you can trust me —I can carry out a simpler liquid biopsy process. Although the liquid biopsy avoid copliacation, common liquid biopsy required laboratory apparatus such as PCR instrument during the detection of ctDNA. In my work, my DNA device and proteins that needed for detection will mixed in a E.coli cell-free system and freeze-dried on a paper screening chip. You will be seeing me in a convenient kit that contains all tools you need for detection, including hemostix, a hand power centrifuge, NASBA reaction system and the paper screening chip.</p>
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<p>Please, click <a href="https://2017.igem.org/Team:BGIC-Union/Project">  here </a> and know more about me as well as the product!</p>
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<p>Please, click <a href="https://2017.igem.org/Team:BGIC-Union/Project">  here </a> and know more about me as well as the product!</p>
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Revision as of 11:22, 3 October 2017

Team：BGIC-Union

Team Gathering

1+1+1+1+1+1+1+1+1+1+1+1>12
Having been through a month of number theory online learning and intense brain storming, the team settled the basic scheme for the project and met up at Shenzhen BGI-college for the first time.

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JUNE, 12

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JUNE, 21

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Hi, I am dCasentry,

a T7—dCas9 multi-output DNA sensor designed to detect ctDNA!

Cancer is a leading cause of death worldwide, accounting for 8.8 million deaths in 2015, and the most common cause of cancer death is lung cancer (1.69 million deaths). Nowadays, multiple lung cancer detection methods are developed, one of them being liquid biopsy. The technique of liquid biopsy is much better than the popular biopsy as it obviates pain and complication incurred to patients by detecting various tumor markup, including short strand circulating tumor DNA (CtDNA) in the blood. However, it still requires laboratory apparatus such as PCR instrument.

Thus our primary task is to devise a more efficient, rapid and convenient lung cancer prognosis method. We construct a paper chip containing powerful biosensor capable of detecting the fusion gene, EML4-ALK, which only exists only in NSCLC (non small cell lung cancer) patients’ blood. Specific targeted drug for this mutation has been produced, which means patients can receive precise treatment as soon as possible when the detection result is positive.

The biosensor, dCasentry, consists of paired dCas9 protein guided by sgRNA and linked with split T7 RNA polymerase that serves as a switch. When the targeting lung cancer mutation gene is detected, visible signal will be released as the T7 RNA polymerase starts transcription. DCasentry not only deals with low ctDNA concentration in blood but also capable of producing various kinds of signal output, such as GFP and RFP.

We adopt freeze-dried paper chip as the vector of the detection system to simplify its operation as well as storage. The DNA device and proteins needed for detection are mixed with cell-free system and freeze-dried on the paper chip. During testing, the blood sample of the patient provides a liquid environment to reactivate the cell-free system. The product will be presented as a kit, containing including hemostix, a hand power centrifuge, NASBA reaction system and the paper screening chip, that can be widely used on the medical field.

Please, click here and know more about me as well as the product!

EXTRACT

CENTRIFUGE

ctDNA AMPLIFICATION

DETECTION

SIGNALING

TEST CHIP