Difference between revisions of "Part:BBa K3791019:Design"
 
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The repeat sequence was extracted from the parts registry (<html><a href="https://parts.igem.org/Part:BBa_K2927006" target="_blank" onmouseover="this.style.color='#ff724a';" onmouseout="this.style.color='#ff9515';" style="color: #ff9515; text-decoration: none;">BBa_K2927006</a></html>), whereas the spacer one was designed with the constraint that it had to be complementary to the sequence of the spectinomycin-resistance gene aiming to be detected (as explained in its own part page: <html><a href="https://parts.igem.org/Part:BBa_K3791004" target="_blank" onmouseover="this.style.color='#ff724a';" onmouseout="this.style.color='#ff9515';" style="color: #ff9515; text-decoration: none;">BBa_K3791004</a></html>).
 
The repeat sequence was extracted from the parts registry (<html><a href="https://parts.igem.org/Part:BBa_K2927006" target="_blank" onmouseover="this.style.color='#ff724a';" onmouseout="this.style.color='#ff9515';" style="color: #ff9515; text-decoration: none;">BBa_K2927006</a></html>), whereas the spacer one was designed with the constraint that it had to be complementary to the sequence of the spectinomycin-resistance gene aiming to be detected (as explained in its own part page: <html><a href="https://parts.igem.org/Part:BBa_K3791004" target="_blank" onmouseover="this.style.color='#ff724a';" onmouseout="this.style.color='#ff9515';" style="color: #ff9515; text-decoration: none;">BBa_K3791004</a></html>).
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===References===
 
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[1] Campa, C. C., Weisbach, N. R., Santinha, A. J., Incarnato, D., & Platt, R. J. (2019). <b>Multiplexed genome engineering by Cas12a and CRISPR arrays encoded on single transcripts.</b> <i>Nature Methods, 16</i>(9), 887–893. <html><a href="https://doi.org/10.1038/s41592-019-0508-6" target="_blank" onmouseover="this.style.color='#ff724a';" onmouseout="this.style.color='#ff9515';" style="color: #ff9515; text-decoration: none;">https://doi.org/10.1038/s41592-019-0508-6</a></html>
 
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<br> [2] Paul, B., & Montoya, G. (2020). <b>CRISPR-Cas12a: Functional overview and applications.</b> <i>Biomedical Journal, 43</i>(1), 8–17. <html><a href="https://doi.org/10.1016/j.bj.2019.10.005" target="_blank" onmouseover="this.style.color='#ff724a';" onmouseout="this.style.color='#ff9515';" style="color: #ff9515; text-decoration: none;">https://doi.org/10.1016/j.bj.2019.10.005</a></html>
 

Latest revision as of 21:10, 21 October 2021


Efficient gRNA Spectinomycin


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

The gRNA architecture includes two DR separated by the spacer with 4 additional base pairs downstream. So the final structure is: repeat + spacer + 4 nucleotides + repeat.


Source

The repeat sequence was extracted from the parts registry (BBa_K2927006), whereas the spacer one was designed with the constraint that it had to be complementary to the sequence of the spectinomycin-resistance gene aiming to be detected (as explained in its own part page: BBa_K3791004).