Expression and differentiation between OCT4A and its Pseudogenes in human ESCs and differentiated adult somatic cells
Authors: Sakthikumar Ambady
Christopher Malcuit
Primoz Rozman
Olga Kashpur
Tanja Dominko
Mojca Jez
Lucy Vilner
Alexandra Grella
Journal: PLoS ONE (2014)
Discipline: Multidisciplinary
Keywords: POU5F1 gene; OCT4 transcription factor; pluripotency; alternative splicing; alternative translation
Affiliations: Blood Transfusion Centre of Slovenia, Ljubljana, Slovenia., Department of Biomedical Engineering, Worcester Polytechnic Institute, Worcester, Massachusetts, United States of America., Department of Biology and Biotechnology, Worcester Polytechnic Institute, Worcester, Massachusetts, United States of America., Bioengineering Institute, Worcester Polytechnic Institute, Worcester, Massachusetts, United States of America ; CellThera, Inc., Worcester, Massachusetts, United States of America., CellThera, Inc., Worcester, Massachusetts, United States of America., Department of Biology and Biotechnology, Worcester Polytechnic Institute, Worcester, Massachusetts, United States of America ; Bioengineering Institute, Worcester Polytechnic Institute, Worcester, Massachusetts, United States of America ; CellThera, Inc., Worcester, Massachusetts, United States of America.

Abstract:  The POU5F1 gene codes for the OCT4 transcription factor, which is one of the key regulators of pluripotency. Its transcription, alternative splicing, and alternative translation leading to the synthesis of the active, nuclear localized OCT4A has been described in detail. Much less, however, is known about actively transcribed OCT4 pseudogenes, several of which display high homology to OCT4A and can be expressed and translated into proteins. Using RT-PCR followed by pseudogene-specific restriction digestion, cloning, and sequencing we discriminate between OCT4A and transcripts for pseudogenes 1, 3 and 4. We show that expression of OCT4 and its pseudogenes follows a developmentally-regulated pattern in differentiating hESCs, indicating a tight regulatory relationship between them. We further demonstrate that differentiated human cells from a variety of tissues express exclusively pseudogenes. Expression of OCT4A can, however be triggered in adult differentiated cells by oxygen and FGF2-dependent mechanisms. 

Retracted: No

PubPeer comments:
 July 2014 correction, which reads:  Due to an error in the preparation of Figure 3, the GAPDH panel for hESCs in Figure 3A is incorrect and duplicates the GAPDH panel for differentiating hESCs in Figure 3B. The authors would also like to provide a clarification in relation to Figure 3C. The bands in the original gel for CRIPTO1 and NANOG were re-arranged to display them in the order d0, d3, d7 in the published figure, instead of the original loading order of d0, d7, d3. The authors are supplying a corrected Figure 3 that includes the correct GAPDH panel for Figure 3A and a revised Figure 3C where the original order of the lanes is maintained. The authors apologize for the inaccuracies in the original Figure 3. These changes do not affect the results and conclusions reported in the article.  Here is the original Figure 3, as reported to the journal in March 2014, with the duplication marked with red boxes. ![file] ,