Coding

Part:BBa_K3894016

Designed by: Jiaqi Zhang   Group: iGEM21_NEFU_China   (2021-10-17)


ZF
Through designing a ZF to bind a specific sequence and attaching it to a single strand-cleavage enzyme, we could generate a guided nickase to create a nick at a desirable site.

Description

To enhance the specificity and programmability of our detection system, we also planned to use a zinc finger (ZF) protein as an anchoring tool. Through designing a ZF to bind a specific sequence and attaching it to a single strand-cleavage enzyme, we could generate a guided nickase to create a nick at a desirable site. This construction integrated the functions of DNA anchorage and nickase cleavage. Individual zinc finger (ZF) domains that recognize DNA triplets with high specificity and affinity can be used to design target DNA sequences that are specific for nearly any region in the genome. These domains can be treated as modular units and assembled to create a polydactyl protein that recognizes extended DNA sequences[1]. The iGEM 2021 NEFU_China also designed ZF to fuse with the BbvCI-R1（BBa_K3894017） enzyme that can create single strand nicks.

Experience

The iGEM 2021 NEFU_China used the Zinc Finger Tools created by Jeffrey G. Mandell,et al. Through exploring the experimental characteristic database of the zinc finger domains, the expected amino acids of a zinc finger protein bound to a selected target can be simulated and predicted.

References

[1] J. Mandell and C. Barbas, Zinc Finger Tools: custom DNA-binding domains for transcription factors and nucleases, Oxford University Press, W516-W523, 2015.

Sequence and Features Sequence and Features


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]


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Categories
Parameters
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