Difference between revisions of "Part:BBa K3402008"
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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. | 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. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
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| + | <i>NLS</i> was connected to the 3' end of the <i>yeGFP</i>([https://parts.igem.org/Part:BBa_K3402000# 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. | ||
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| + | [[Image:Without NLS.png|500px|thumb|center|Without the <i>NLS</i> (nuclear localization signal peptide)]] | ||
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| + | [[Image:With NLS.png|500px|thumb|center|With the <i>NLS</i> (nuclear localization signal peptide)]] | ||
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Revision as of 05:57, 11 October 2020
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.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]