Coding

Part:BBa_K1150003

Designed by: Freiburg 2013   Group: iGEM13_Freiburg   (2013-09-16)
Revision as of 16:29, 3 October 2013 by Lisaschmunk (Talk | contribs)

G9a

G9a
Function Histone Methyltransferase
Use in Mammalian cells
RFC standard RFC 25
Backbone pSB1C3
Organism Mus musculus
Source Albert Jeltsch, Stuttgart
Submitted by [http://2013.igem.org/Team:Freiburg Freiburg 2013]

This part contains the catalytically active set domain of G9a on the murine EHMT2 gene. G9a is able to transfer methyl groups to lysine 9 of histone 3 (H3K9), which is part of every nucleosome. In regions of open chromatin structures H3K9 methylation can lead to chromatin condensation and thereby inactivation of the concerned locus. This G9a part alone contains the histone methyltransferase domain needed for transcriptional repression and can be combined with dCas9 Part:BBa_K1150000 to target different loci for transcriptional repression of gene expression.

Usage and Biology

This part belongs to the uniCAS toolkit of iGEM Team Freiburg 2013. In fusion with dCas9 Part:BBa_K1150024 this part can be used to effectively repress target genes by di- or tri-methylation of H3K9. We offer four devices using this part: - the uniCAS Histone Modifier (CMV promoter) Part:BBa_K1150024 favorite part! - the uniCAS Histone Modifier (SV40 promoter) Part:BBa_K1150023 uses a medium strong SV40 promoter to optimize this device for means were a strong expression is suboptimal. - the uniCAS Red Light Switch Part II - Histone Modifier Part:BBa_K1150028 is a result of combining G9a with part of our red light switch, Phytochrome BPart:BBa_K1150004. PhyB will dimerize with PIF6 upon red light stimulus. PIF6 is fused to dCas9 in the uniCAS Red Light Switch Part I - Stimulator Part:BBa_K1150025 . Using these two devices G9a is brought into proximity of the DNA targeting dCas9 and can therefore site specifically transfer methyl groups and repress gene expression. - the uniCAS UV Light Switch Part II - Histone Modifier Part:BBa_K1150032


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 597
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None