Coding

Part:BBa_K3402008

Designed by: Yibo Shi   Group: iGEM20_Jiangnan_China   (2020-10-01)
Revision as of 05:57, 11 October 2020 by ZZ 0991 (Talk | contribs)


NLS

Because of the high molecular weight of Cas9 nuclease which cannot freely shuttle through the nuclear pores, we have to add a nuclear localization sequence(NLS) to guide it through the nucleus.

Usage and Biology

NLS was connected to the 3' end of the yeGFP(BBa_K3402000) gene to construct the plasmid, then we transferred the plasmid to observe the distribution of green fluorescence. If the fluorescent region was concentrated in the nucleus, this sequence could be used for the subsequent establishment of the CRISPR/Cas9 system.

Without the NLS (nuclear localization signal peptide)
With the NLS (nuclear localization signal peptide)

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]


[edit]
Categories
Parameters
None