Difference between revisions of "Part:BBa K4245207:Design"

 
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<partinfo>BBa_K4245207 short</partinfo>
 
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===Design Notes===
 
===Design Notes===
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hsa-miR-208a-3p RCA Padlock Probe
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<br>
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A padlock probe, which is often 30-150 nucleotides in length, is a single-stranded DNA sequence designed to recognize a specific target sequence. The “arms” of a padlock probe are the ends of the ssDNA that are complementary to a specific target sequence. The middle sequence (the sequence between the arms) can be specifically designed to perform a function once amplified. (Nilsson et al., 1994)
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<br>
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<br>
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Lambert iGEM found three specific miRNAs — hsa-miR-1-3p (<partinfo>BBa_K4245006</partinfo>), hsa-mir-133a-3p (<partinfo>BBa_K4245009</partinfo>), and hsa-miR-208a-3p (<partinfo>BBa_K4245011</partinfo>)— to be upregulated in correlation to CAD (Kaur et al., 2020). For miRNA 208a-3p, we designed two complementary arms, <partinfo>BBa_K4245105</partinfo>, the 3' arm for hsa-miR-208a-3p and <partinfo>BBa_K4245112</partinfo>, the 5' arm for hsa-miR-208a-3p. For the reporter, we decided on both <partinfo>BBa_K4245130</partinfo>/ <partinfo>BBa_K4245132</partinfo> the FAM and BHQ1 labeled linear probes, and <partinfo>BBa_K4245134</partinfo>/ <partinfo>BBa_K4245135</partinfo>, the DNA fluorescent aptamer split lettuce, due to their frugality and similar wavelength to GFP. 
  
 
===Source===
 
===Source===
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===References===
 
===References===
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Kaur, A., Mackin, S. T., Schlosser, K., Wong, F. L., Elharram, M., Delles, C., Stewart, D. J., Dayan, N., Landry, T., & Pilote, L. (2019). Systematic review of microrna biomarkers in acute coronary syndrome and stable coronary artery disease. Cardiovascular Research, 116(6), 1113–1124. https://doi.org/10.1093/cvr/cvz302
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<br>
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Nilsson, M., Malmgren, H., Samiotaki, M., Kwiatkowski, M., Chowdhary, B. P., & Landegren, U. (1994). Padlock probes: Circularizing oligonucleotides for localized DNA detection. Science, 265(5181), 2085–2088. https://doi.org/10.1126/science.7522346

Revision as of 22:14, 11 October 2022

hsa-miR-208a-3p RCA Padlock Probe


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes


A padlock probe, which is often 30-150 nucleotides in length, is a single-stranded DNA sequence designed to recognize a specific target sequence. The “arms” of a padlock probe are the ends of the ssDNA that are complementary to a specific target sequence. The middle sequence (the sequence between the arms) can be specifically designed to perform a function once amplified. (Nilsson et al., 1994)

Lambert iGEM found three specific miRNAs — hsa-miR-1-3p (BBa_K4245006), hsa-mir-133a-3p (BBa_K4245009), and hsa-miR-208a-3p (BBa_K4245011)— to be upregulated in correlation to CAD (Kaur et al., 2020). For miRNA 208a-3p, we designed two complementary arms, BBa_K4245105, the 3' arm for hsa-miR-208a-3p and BBa_K4245112, the 5' arm for hsa-miR-208a-3p. For the reporter, we decided on both BBa_K4245130/ BBa_K4245132 the FAM and BHQ1 labeled linear probes, and BBa_K4245134/ BBa_K4245135, the DNA fluorescent aptamer split lettuce, due to their frugality and similar wavelength to GFP.

Source

hsa-miR-208a-3p RCA Padlock Probe

References

Kaur, A., Mackin, S. T., Schlosser, K., Wong, F. L., Elharram, M., Delles, C., Stewart, D. J., Dayan, N., Landry, T., & Pilote, L. (2019). Systematic review of microrna biomarkers in acute coronary syndrome and stable coronary artery disease. Cardiovascular Research, 116(6), 1113–1124. https://doi.org/10.1093/cvr/cvz302
Nilsson, M., Malmgren, H., Samiotaki, M., Kwiatkowski, M., Chowdhary, B. P., & Landegren, U. (1994). Padlock probes: Circularizing oligonucleotides for localized DNA detection. Science, 265(5181), 2085–2088. https://doi.org/10.1126/science.7522346