Qiuxinyuan12 (Talk | contribs) |
Qiuxinyuan12 (Talk | contribs) |
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<h2> | <h2> | ||
− | <span>Abstract</span> | + | <span>Abstract</span> |
</h2> | </h2> | ||
+ | <p> | ||
+ | <b><span style="font-size:18px;"> </span></b> | ||
+ | </p> | ||
<p> | <p> | ||
<span style="font-size:18px;">MicroRNAs, serve as critical gene | <span style="font-size:18px;">MicroRNAs, serve as critical gene | ||
Line 405: | Line 408: | ||
establishment and further development might provide a new approach for rapid | establishment and further development might provide a new approach for rapid | ||
and low-cost cancer screening, virus detection and curative efficacy | and low-cost cancer screening, virus detection and curative efficacy | ||
− | assessment.</span> | + | assessment.</span> |
− | </p></ | + | </p> |
+ | <p> | ||
+ | <span style="font-size:18px;"> </span> | ||
+ | </p> | ||
<h2> | <h2> | ||
− | <span>Introduction</span> | + | <span>Introduction</span> |
</h2> | </h2> | ||
+ | <p> | ||
+ | <b><span style="font-size:18px;"> </span></b> | ||
+ | </p> | ||
<p> | <p> | ||
<span style="font-size:18px;">Nowadays, cancers, due to their high | <span style="font-size:18px;">Nowadays, cancers, due to their high | ||
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Such situation highlighted the undiminished importance of the development of a | Such situation highlighted the undiminished importance of the development of a | ||
low-cost, easily accessible and rapid tool for early screening and detection of | low-cost, easily accessible and rapid tool for early screening and detection of | ||
− | cancers.</span> </ | + | cancers.</span> |
+ | </p> | ||
+ | <p> | ||
+ | <span style="font-size:18px;"> </span> | ||
+ | </p> | ||
+ | <p> | ||
+ | <b><span style="font-size:14px;">Figure | ||
+ | 1. Global distribution of estimated age-standardized world cancer incidence | ||
+ | rate (ASR) per 100 000 in (A) men, (B) women, and mortality rate (ASR) per 100 | ||
+ | 000 in (C) men and (D) women. (WHO, World cancer report, 2014)</span></b> | ||
</p> | </p> | ||
<p align="center" style="text-align:center;"> | <p align="center" style="text-align:center;"> | ||
− | <b><span style="font-size:18px;"> | + | <b><span style="font-size:18px;"> </span></b> |
− | </p | + | </p> |
<p> | <p> | ||
<span style="font-size:18px;">MicroRNAs (miRNAs), as a kind of small | <span style="font-size:18px;">MicroRNAs (miRNAs), as a kind of small | ||
Line 455: | Line 473: | ||
requirements of sensitivity and specificity. At the main time, such method | requirements of sensitivity and specificity. At the main time, such method | ||
still relies on equipment such as Fluoresce microplate readers or fluorescence | still relies on equipment such as Fluoresce microplate readers or fluorescence | ||
− | microscopes, which are highly costly.</span> | + | microscopes, which are highly costly.</span> |
</p> | </p> | ||
− | <p | + | <p> |
− | <b><span style="font-size:18px;"> | + | <span style="font-size:18px;"> </span> |
+ | </p> | ||
+ | <p> | ||
+ | <b><span style="font-size:14px;">Figure | ||
+ | 2. Workflow for our CRISPR-based blood-microRNA detection system.</span></b> | ||
+ | </p> | ||
+ | <p> | ||
+ | <span style="font-size:14px;">Using sequence information from online | ||
+ | databases, probes for RCA reaction were designed in silico. Once synthesized | ||
+ | and sealed to form the dumbbell structure, the probe, together with other | ||
+ | necessary materials can be embedded into tubes and freeze-dried to remain | ||
+ | stable in room temperature for a relatively long time. For the detection | ||
+ | process, serum samples were pre-treated by boiling in 95℃ for 15 min to expose the | ||
+ | miRNAs completely. The amount of the specific RNA was indicated by a color | ||
+ | difference in the tube from colorless to blue</span> | ||
+ | </p> | ||
+ | <p> | ||
+ | <b><span style="font-size:18px;"> </span></b> | ||
</p> | </p> | ||
<p> | <p> | ||
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to reliably expose miRNAs in serum samples and demonstrated robust detection | to reliably expose miRNAs in serum samples and demonstrated robust detection | ||
with this scheme to compare let-7a concentrations among blood samples collected | with this scheme to compare let-7a concentrations among blood samples collected | ||
− | from NSCLC patients and healthy volunteers.</span> | + | from NSCLC patients and healthy volunteers.</span> |
− | </p></ | + | </p> |
+ | <p> | ||
+ | <span style="font-size:18px;"> </span> | ||
+ | </p> | ||
<h2> | <h2> | ||
<span>Improvements | <span>Improvements | ||
− | we made</span> | + | we made</span> |
</h2> | </h2> | ||
<p> | <p> | ||
− | <span style="font-size:18px;">BBa_K1789003 and BBa_K1789004</span> | + | <b><span style="font-size:18px;"> </span></b> |
+ | </p> | ||
+ | <p> | ||
+ | <span style="font-size:18px;"><strong>BBa_K1789003 and BBa_K1789004</strong></span> | ||
</p> | </p> | ||
<p> | <p> | ||
Line 491: | Line 532: | ||
classical PPI indicator, split-GFP system, developed previously in our project | classical PPI indicator, split-GFP system, developed previously in our project | ||
in iGEM2015 (BBa_K1789003 and BBa_K1789004), was also included in our kit. | in iGEM2015 (BBa_K1789003 and BBa_K1789004), was also included in our kit. | ||
− | Several improvements has been made for this system including:</span> | + | Several improvements has been made for this system including:</span> |
</p> | </p> | ||
<p style="text-indent:-18pt;"> | <p style="text-indent:-18pt;"> | ||
<span style="font-size:18px;">1. Improved | <span style="font-size:18px;">1. Improved | ||
− | characterization for previous parts</span> | + | characterization for previous parts</span> |
</p> | </p> | ||
<p> | <p> | ||
Line 508: | Line 549: | ||
detected by a microplate reader after an overnight culture under various | detected by a microplate reader after an overnight culture under various | ||
conditions. Relative fluorescence | conditions. Relative fluorescence | ||
− | intensity was then calculated with normalization of OD</span><sub><span style="font-size:18px;">600</span></sub><span style="font-size:18px;"> value.</span> | + | intensity was then calculated with normalization of OD</span><sub><span style="font-size:18px;">600</span></sub><span style="font-size:18px;"> value.</span> |
</p> | </p> | ||
<p> | <p> | ||
<span style="font-size:18px;">Results shown a better | <span style="font-size:18px;">Results shown a better | ||
− | SNR under 20℃ | + | SNR under 20℃ and 0.5mM IPTG induction (Figure 3B). Thus |
− | and 0.5mM IPTG induction (Figure 3B). Thus | + | indicating that better performance of such system could be expected under lower |
− | that better performance of such system could be expected under lower | + | culturing temperature.</span> |
− | temperature.</span> | + | </p> |
+ | <p> | ||
+ | <span style="font-size:18px;"> </span> | ||
</p> | </p> | ||
<p align="center" style="text-align:center;"> | <p align="center" style="text-align:center;"> | ||
− | <b><span style="font-size: | + | <b><span style="font-size:14px;">(Figure 3)</span></b> |
+ | </p> | ||
+ | <p> | ||
+ | <span style="font-size:18px;"> </span> | ||
+ | </p> | ||
+ | <p> | ||
+ | <span style="font-size:18px;"> </span> | ||
</p> | </p> | ||
<p style="text-indent:-18pt;"> | <p style="text-indent:-18pt;"> | ||
Line 527: | Line 576: | ||
194-212), GFP11 (residues 213-233) and GFP 1-9 (residues 1-193) [sci-reps 10.103/srep02854]. Due to their short | 194-212), GFP11 (residues 213-233) and GFP 1-9 (residues 1-193) [sci-reps 10.103/srep02854]. Due to their short | ||
length, two small fragments can be easily fused onto proteins with less | length, two small fragments can be easily fused onto proteins with less | ||
− | affection on their folding (figure 4A).</span> | + | affection on their folding (figure 4A).</span> |
+ | </p> | ||
+ | <p> | ||
+ | <span style="font-size:18px;"> </span> | ||
</p> | </p> | ||
<p align="center" style="text-align:center;"> | <p align="center" style="text-align:center;"> | ||
− | <b><span style="font-size: | + | <b><span style="font-size:14px;">(Figure 4)</span></b> |
+ | </p> | ||
+ | <p align="center" style="text-align:center;"> | ||
+ | <b><span style="font-size:18px;"> </span></b> | ||
</p> | </p> | ||
<p> | <p> | ||
<span style="font-size:18px;">Comparing with previous | <span style="font-size:18px;">Comparing with previous | ||
split-GFP system, higher SNR was reached under the same expression condition, | split-GFP system, higher SNR was reached under the same expression condition, | ||
− | while the total signal intensity suffered tolerable decrease (Figure 4B).</span> | + | while the total signal intensity suffered tolerable decrease (Figure 4B).</span> |
</p> | </p> | ||
<p> | <p> | ||
− | <span style="font-size:18px;"> </span> | + | <span style="font-size:18px;"> </span> |
</p> | </p> | ||
<h2> | <h2> | ||
− | <span> | + | <span>Reference</span> |
</h2> | </h2> | ||
− | < | + | </br> |
− | <span style="font-size:18px;">1 Jeong, K. E. & Cairns, J. A. Review of economic evidence in the prevention and early detection of colorectal cancer. </span><i><span style="font-size:18px;">Health Econ Rev</span></i> <b><span style="font-size:18px;">3</span></b><span style="font-size:18px;">, 20, doi:10.1186/2191-1991-3-20 (2013).</span> | + | |
− | </ | + | </p> |
− | < | + | <p style="text-indent:-36pt;"> |
− | <span style="font-size:18px;">2 Etzioni, R.</span><i><span style="font-size:18px;"> et al.</span></i><span style="font-size:18px;"> The case for early detection. </span><i><span style="font-size:18px;">Nat Rev Cancer</span></i> <b><span style="font-size:18px;">3</span></b><span style="font-size:18px;">, 243-252, doi:10.1038/nrc1041 (2003).</span> | + | <span style="font-size:18px;">1 Jeong, |
− | </ | + | K. E. & Cairns, J. A. Review of economic evidence in the prevention and |
− | < | + | early detection of colorectal cancer. </span><i><span style="font-size:18px;">Health |
− | <span style="font-size:18px;">3 Wolf, A. M.</span><i><span style="font-size:18px;"> et al.</span></i><span style="font-size:18px;"> American Cancer Society guideline for the early detection of prostate cancer: update 2010. </span><i><span style="font-size:18px;">CA Cancer J Clin</span></i> <b><span style="font-size:18px;">60</span></b><span style="font-size:18px;">, 70-98, doi:10.3322/caac.20066 (2010).</span> | + | Econ Rev</span></i> <b><span style="font-size:18px;">3</span></b><span style="font-size:18px;">, 20, |
− | </ | + | doi:10.1186/2191-1991-3-20 (2013).</span> |
− | < | + | </p> |
− | <span style="font-size:18px;">4 McPhail, S., Johnson, S., Greenberg, D., Peake, M. & Rous, B. Stage at diagnosis and early mortality from cancer in England. </span><i><span style="font-size:18px;">Br J Cancer</span></i> <b><span style="font-size:18px;">112 Suppl 1</span></b><span style="font-size:18px;">, S108-115, doi:10.1038/bjc.2015.49 (2015).</span> | + | <p style="text-indent:-36pt;"> |
− | </ | + | <span style="font-size:18px;">2 Etzioni, R.</span><i><span style="font-size:18px;"> et al.</span></i><span style="font-size:18px;"> The case for early detection. </span><i><span style="font-size:18px;">Nat Rev Cancer</span></i> <b><span style="font-size:18px;">3</span></b><span style="font-size:18px;">, |
− | < | + | 243-252, doi:10.1038/nrc1041 (2003).</span> |
− | <span style="font-size:18px;">5 Ambros, V. MicroRNA pathways in flies and worms: growth, death, fat, stress, and timing. </span><i><span style="font-size:18px;">Cell</span></i> <b><span style="font-size:18px;">113</span></b><span style="font-size:18px;">, 673-676 (2003).</span> | + | </p> |
− | </ | + | <p style="text-indent:-36pt;"> |
− | < | + | <span style="font-size:18px;">3 Wolf, A. M.</span><i><span style="font-size:18px;"> et al.</span></i><span style="font-size:18px;"> American Cancer Society guideline for the early detection |
− | <span style="font-size:18px;">6 Lu, J.</span><i><span style="font-size:18px;"> et al.</span></i><span style="font-size:18px;"> MicroRNA expression profiles classify human cancers. </span><i><span style="font-size:18px;">Nature</span></i> <b><span style="font-size:18px;">435</span></b><span style="font-size:18px;">, 834-838, doi:10.1038/nature03702 (2005).</span> | + | of prostate cancer: update 2010. </span><i><span style="font-size:18px;">CA |
− | </ | + | Cancer J Clin</span></i> <b><span style="font-size:18px;">60</span></b><span style="font-size:18px;">, 70-98, |
− | < | + | doi:10.3322/caac.20066 (2010).</span> |
− | <span style="font-size:18px;">7 Jansson, M. D. & Lund, A. H. MicroRNA and cancer. </span><i><span style="font-size:18px;">Mol Oncol</span></i> <b><span style="font-size:18px;">6</span></b><span style="font-size:18px;">, 590-610, doi:10.1016/j.molonc.2012.09.006 (2012).</span> | + | </p> |
− | </ | + | <p style="text-indent:-36pt;"> |
− | < | + | <span style="font-size:18px;">4 McPhail, S., Johnson, S., Greenberg, D., |
− | <span style="font-size:18px;">8 Schultz, N. A.</span><i><span style="font-size:18px;"> et al.</span></i><span style="font-size:18px;"> MicroRNA biomarkers in whole blood for detection of pancreatic cancer. </span><i><span style="font-size:18px;">JAMA</span></i> <b><span style="font-size:18px;">311</span></b><span style="font-size:18px;">, 392-404, doi:10.1001/jama.2013.284664 (2014).</span> | + | Peake, M. & Rous, B. Stage at diagnosis and early mortality from cancer in |
− | </ | + | England. </span><i><span style="font-size:18px;">Br J Cancer</span></i> <b><span style="font-size:18px;">112 Suppl 1</span></b><span style="font-size:18px;">, S108-115, |
− | < | + | doi:10.1038/bjc.2015.49 (2015).</span> |
− | <span style="font-size:18px;">9 Chen, X.</span><i><span style="font-size:18px;"> et al.</span></i><span style="font-size:18px;"> Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. </span><i><span style="font-size:18px;">Cell Res</span></i> <b><span style="font-size:18px;">18</span></b><span style="font-size:18px;">, 997-1006, doi:10.1038/cr.2008.282 (2008).</span> | + | </p> |
− | </ | + | <p style="text-indent:-36pt;"> |
− | < | + | <span style="font-size:18px;">5 Ambros, V. MicroRNA pathways in flies and |
− | <span style="font-size:18px;">10 Hunt, E. A., Broyles, D., Head, T. & Deo, S. K. MicroRNA Detection: Current Technology and Research Strategies. </span><i><span style="font-size:18px;">Annu Rev Anal Chem (Palo Alto Calif)</span></i> <b><span style="font-size:18px;">8</span></b><span style="font-size:18px;">, 217-237, doi:10.1146/annurev-anchem-071114-040343 (2015).</span> | + | worms: growth, death, fat, stress, and timing. </span><i><span style="font-size:18px;">Cell</span></i> <b><span style="font-size:18px;">113</span></b><span style="font-size:18px;">, 673-676 |
− | </ | + | (2003).</span> |
− | < | + | </p> |
− | <span style="font-size:18px;">11 Cissell, K. A., Rahimi, Y., Shrestha, S., Hunt, E. A. & Deo, S. K. Bioluminescence-based detection of microRNA, miR21 in breast cancer cells. </span><i><span style="font-size:18px;">Anal Chem</span></i> <b><span style="font-size:18px;">80</span></b><span style="font-size:18px;">, 2319-2325, doi:10.1021/ac702577a (2008).</span> | + | <p style="text-indent:-36pt;"> |
− | </ | + | <span style="font-size:18px;">6 Lu, J.</span><i><span style="font-size:18px;"> et al.</span></i><span style="font-size:18px;"> MicroRNA expression profiles classify human cancers. </span><i><span style="font-size:18px;">Nature</span></i> <b><span style="font-size:18px;">435</span></b><span style="font-size:18px;">, 834-838, doi:10.1038/nature03702 (2005).</span> |
− | < | + | </p> |
− | <span style="font-size:18px;">12 Venkatesan, B. M. & Bashir, R. Nanopore sensors for nucleic acid analysis. </span><i><span style="font-size:18px;">Nature nanotechnology</span></i> <b><span style="font-size:18px;">6</span></b><span style="font-size:18px;">, 615-624, doi:10.1038/nnano.2011.129 (2011).</span> | + | <p style="text-indent:-36pt;"> |
− | </ | + | <span style="font-size:18px;">7 Jansson, M. D. & Lund, A. H. MicroRNA |
− | < | + | and cancer. </span><i><span style="font-size:18px;">Mol Oncol</span></i> <b><span style="font-size:18px;">6</span></b><span style="font-size:18px;">, 590-610, |
− | <span style="font-size:18px;">13 Deng, R.</span><i><span style="font-size:18px;"> et al.</span></i><span style="font-size:18px;"> Toehold-initiated rolling circle amplification for visualizing individual microRNAs in situ in single cells. </span><i><span style="font-size:18px;">Angew Chem Int Ed Engl</span></i> <b><span style="font-size:18px;">53</span></b><span style="font-size:18px;">, 2389-2393, doi:10.1002/anie.201309388 (2014).</span> | + | doi:10.1016/j.molonc.2012.09.006 (2012).</span> |
− | </ | + | </p> |
+ | <p style="text-indent:-36pt;"> | ||
+ | <span style="font-size:18px;">8 Schultz, N. A.</span><i><span style="font-size:18px;"> et al.</span></i><span style="font-size:18px;"> MicroRNA biomarkers in whole blood for detection of | ||
+ | pancreatic cancer. </span><i><span style="font-size:18px;">JAMA</span></i> <b><span style="font-size:18px;">311</span></b><span style="font-size:18px;">, 392-404, | ||
+ | doi:10.1001/jama.2013.284664 (2014).</span> | ||
+ | </p> | ||
+ | <p style="text-indent:-36pt;"> | ||
+ | <span style="font-size:18px;">9 Chen, X.</span><i><span style="font-size:18px;"> et al.</span></i><span style="font-size:18px;"> Characterization of microRNAs in serum: a novel class of | ||
+ | biomarkers for diagnosis of cancer and other diseases. </span><i><span style="font-size:18px;">Cell Res</span></i> <b><span style="font-size:18px;">18</span></b><span style="font-size:18px;">, 997-1006, | ||
+ | doi:10.1038/cr.2008.282 (2008).</span> | ||
+ | </p> | ||
+ | <p style="text-indent:-36pt;"> | ||
+ | <span style="font-size:18px;">10 Hunt, E. A., Broyles, D., Head, T. & | ||
+ | Deo, S. K. MicroRNA Detection: Current Technology and Research Strategies. </span><i><span style="font-size:18px;">Annu Rev Anal Chem (Palo Alto Calif)</span></i> <b><span style="font-size:18px;">8</span></b><span style="font-size:18px;">, 217-237, doi:10.1146/annurev-anchem-071114-040343 | ||
+ | (2015).</span> | ||
+ | </p> | ||
+ | <p style="text-indent:-36pt;"> | ||
+ | <span style="font-size:18px;">11 Cissell, K. A., Rahimi, Y., Shrestha, S., | ||
+ | Hunt, E. A. & Deo, S. K. Bioluminescence-based detection of microRNA, miR21 | ||
+ | in breast cancer cells. </span><i><span style="font-size:18px;">Anal Chem</span></i> <b><span style="font-size:18px;">80</span></b><span style="font-size:18px;">, 2319-2325, doi:10.1021/ac702577a | ||
+ | (2008).</span> | ||
+ | </p> | ||
+ | <p style="text-indent:-36pt;"> | ||
+ | <span style="font-size:18px;">12 Venkatesan, B. M. & Bashir, R. Nanopore | ||
+ | sensors for nucleic acid analysis. </span><i><span style="font-size:18px;">Nature | ||
+ | nanotechnology</span></i> <b><span style="font-size:18px;">6</span></b><span style="font-size:18px;">, 615-624, | ||
+ | doi:10.1038/nnano.2011.129 (2011).</span> | ||
+ | </p> | ||
+ | <p style="text-indent:-36pt;"> | ||
+ | <span style="font-size:18px;">13 Deng, R.</span><i><span style="font-size:18px;"> et al.</span></i><span style="font-size:18px;"> Toehold-initiated rolling circle amplification for visualizing | ||
+ | individual microRNAs in situ in single cells. </span><i><span style="font-size:18px;">Angew Chem Int Ed Engl</span></i> <b><span style="font-size:18px;">53</span></b><span style="font-size:18px;">, | ||
+ | 2389-2393, doi:10.1002/anie.201309388 (2014).</span> | ||
+ | </p> | ||
<p> | <p> | ||
− | <span style="font-size:18px;"> </span> | + | <span style="font-size:18px;"> </span> |
</p> | </p> | ||
− | |||
Revision as of 17:13, 13 October 2016