TP53 is the most intensely researched gene in human genome. Despite of decades of enormous effort, there are many unanswered questions and understudied fields the TP53 biology. TP53 gene encodes a remarkable transcription factor p53 which directly transactivates about one hundred of conserved “core response” genes involved in multiple tumor-suppressive programs including apoptosis cell cycle arrest, DNA repair, or senescence. However, there is a group of genes undoubtedly regulated by p53 with poorly defined role in carcinogenesis and cellular response to p53 activation. One of such understudied p53 targets is KCNN4, a potassium calcium-activated channel gene.  
 
Rationale
Besides of several dozens of p53 target genes with well-defined function, many genes within this network represent a grey area of knowledge. Understanding better the role of those genes is required for elucidating tumor-suppressive mechanisms driven by p53. Importantly, as p53 activity is context-dependent with distinct characteristic influenced by differentiation, knowing more about p53 target genes is essential for defining roles of p53 across various tissues.    
 
Hypothesis
KCNN4 is induced by p53 activity in colorectal carcinoma lines on the protein level. 
 
Specific Aims
To test the hypothesis, we will pursuit following Specific Aims: 
Aim 1. To identify p53 RE in the KCNN4 locus. 
Aim 2. To verify KCNN4 induction on protein level by western blot.
Aim 3. To validate KCNN4 antibody specificity by KCNN4 suppression by shRNA.