We study the seam cells of C.
elegans because they divide just
like stem cells in humans
- C. elegans is cheap
- C. elegans grow fast
We are going to mainly study to cell division in V1-V4
- First there is an asymmetrical division – L1
- Then the posterior daughter has a symmetrical
division and after that an asymmetrical division – L2
- That is followed by an asymmetrical division – L3
- And lastly there is another asymmetrical division
– L4
- After the L4 stage, all the seam cells have differentiated
Research Question: What other genes are involved in the regulation of the balance
between seam cell proliferation and differentiation?
For this research we use a C. elegans Wildtype (JR667) and a C. elegans tumour-like
phenotype, which leaks Unc-62 (AW673)
- Unc-62 is a transcription factor which regulates the transcription (DNA to RNA) in the
nucleus
- General factors bind to the promotor region, the TATA-box (GTF)
Transcription factors could be either:
- An activator
- An inhibitor/repressor
Activators
- Up-regulation of expression
- By mediating complex formation
Enhancer sequence
- 50 – 1500 bp in size
- 10 – 1000 kb distant from gene
, Unc-62 --- gene 16 seam cells
Unc-62 --- gene hyperplasia of seam cells
When your gene is not a regulator, it will not play
a part in the lab
- Binding to the DNA is a requirement for a
transcription factor
ChIP – Chromatin ImmunoPrecipitation assay
- Lysis of the cell
- Antibodies against Unc-62
- Every sequence to which it was bound, is known Wormbase
modENCODE for Unc-62 – Model Organism
Encyclopeadia of DNA elements
- Pulls out 31029 binding sites, but there are some repeats of the same genes
- Filter for 2.5 kb up- and downstream and within a gene for binding sites
- Then manually look and select those which might be of interest
Final list of 7358 genes
Slowly working through them
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