Coding

Part:BBa_K1150002

Designed by: Freiburg 2013   Group: iGEM13_Freiburg   (2013-09-16)
Revision as of 23:29, 3 October 2013 by ThomasJ (Talk | contribs)

KRAB

KRAB
Function Repressor domain
Use in Mammalian cells
Organism Homo sapiens
RFC standard RFC 25
Backbone pSB1C3
Source Konrad Müller
Submitted by [http://2013.igem.org/Team:Freiburg Freiburg 2013]
Fig. 1 Structural Mechanism of KRAB, as suggested Urrutia, R., 2003.

Krüppel-associated Box repressor domains - commonly termed as KRAB - are highly conserved polypeptide motifs and were first functionally characterized in 1991 (Rosati et al., 1991). As they constitute about one third of all human zinc finger transcription factors, key regulatory features in higher eukaryotic transcrip-
tomics are suggested (Witzgall et al., 1994). Even in terms of tetrapod evolution, the role of their great abundance has been extensively discussed (Birtle, 2006). Even though KRAB minimal domains are usually no longer than 50-75 amino acids, their mechanism of function remains complex.


Common biochemical models suggest a key role in epigenetic silencing, by recruiting a scaffold of diverse proteins - amongst others histone deacetylases and histone methyltransferases (Urrutia, 2003). Til date in 2013, KRAB repressor domains were attached to several DNA binding proteins such as tetR, TAL effec-
tors and [http://2013.igem.org/Team:Freiburg/Project/effector#repression dCas9] - thereby efficiently silencing gene expression downstream of desired target promoters.

The latter combinatorial attempt was conducted twice by Team Freiburg 2013, which resulted in two composite parts - SV40-dCas9-KRAB and CMV-dCas9-KRAB.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 244
    Illegal SapI.rc site found at 208


References

Rosati, M. et al. (1991). Members of the zinc finger protein gene family sharing a conserved N-terminal module. Nucleic acids research 19, 5661-5667.
Witzgall, R. et al. (1994). The Krüppel-associated box-A domain of zinc finger proteins mediates transcriptional repression. Proc Nati Acad Sci 91, 4514-4518.
Birtle, Z. and Ponting, C. (2006). Meisetz and the birth of the KRAB motif. Bioinformatics 22, 2841-2845.
Urrutia, R. (2003). KRAB-containing zinc-finger repressor proteins. Genome Biology 4, 4:231.

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