Difference between revisions of "Part:BBa K3791010:Design"
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===References===
 
===References===
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[1] New England Biolabs. (s/f). <b>How do I design a guide RNA for use with EnGen Lba Cas12a?</b> Neb.com. From: https://international.neb.com/faqs/2018/05/03/how-do-i-design-a-guide-rna-for-use-with-engen-lba-cas12a
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[1] Nguyen, L. T., Smith, B. M., & Jain, P. K. (2020). Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection. Nature Communications, 11(1). https://doi.org/10.1038/s41467-020-18615-1

Revision as of 20:53, 21 October 2021


gRNA Ampicillin construct


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 repeat sequence must be placed before the spacer sequence, that is, in the 5’ extreme. It is also required to have a length between 18 and 24 nucleotides, in this case adjusting it to a consensus G-C content of approximately 50%.

Source

The repeat and the T7 promoter sequences were extracted from the parts registry (BBa_K2927006) (BBa_K1614000), whereas the spacer one was designed with the constraint that it had to be complementary to the sequence of the ampicillin-resistance gene aiming to be detected (as explained in its own part page: BBa_K3791000).

References

[1] Nguyen, L. T., Smith, B. M., & Jain, P. K. (2020). Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection. Nature Communications, 11(1). https://doi.org/10.1038/s41467-020-18615-1