DNA methylation is a crucial part of normal organismal development and cellular differentiation in higher organisms. DNA methylation stably alters the gene expression pattern in cells such that cells can "remember where they have been" or decrease gene expression; for example, cells programmed to be pancreatic islets during embryonic development remain pancreatic islets throughout the life of the organism without continuing signals telling them that they need to remain islets.
By now it is clear that Dnmt1, the major eukaryotic DNA methyltransferase, faithfully maintains genome-wide methylation patterns and plays an essential role in the epigenetic network controlling gene expression and genome stability during development.
The new Dnmt1-Chromobody® comprises the antigen binding domain (VHH) derived from Alpaca heavy chain antibodies genetically fused to TagRFP or TagGFP2 (Evrogen).
Dnmt1-Chromobody® in live cells
HeLa cells were transfected with the Dnmt1-Chromobody® plasmid and imaged in live imaging medium: DME w/o phenol red, gentam 1:1000, glutamin 1:100, FCS 5%.
The cells were imaged every 30 minutes for 24 hours.
In G phase cells the signal is homogeneously distributed through the nucleus and cytoplasm. Over time during S phase granules appear in the nucleus as replication foci form, finally the granularity disappears and the cells divide.
Still images of Dnmt1-Chromobody® transfected HeLa cells throughout the cell cycle. Again, formation of replication foci can be seen, as the cells progress normally as the Dnmt1-Chromobody® does not affect normal cell function.
Use the Dnmt1-Chromobody® to:
- Trace dynamic changes during the cell cycle in real time
- Monitor the distribution of an endogenous cell cycle marker protein - unlike with fluorescent fusion proteins there are no overexpression artifacts or cytotoxic effects
Only for research applications, not for diagnostic or therapeutic use.