Targeted disruption of the GAS41 gene encoding a putative transcription factor indicates that GAS41 is essential for cell viability.
The glioma-amplified sequence (GAS) 41 protein has been proposed to be a transcription factor. To investigate its functional role in vivo, we attempted to knock out the GAS41 gene by targeted disruption in the chicken pre-lymphoid cell line DT40. Heterozygous GAS41+/- cell lines generated by the first round of homologous recombination express approximately half the normal level of GAS41 mRNA. However, a homozygous GAS41-/- cell line with both GAS41 alleles disrupted was not obtained following the second round of transfection, indicating that the GAS41 gene is essential for cell viability. Indeed, homozygous GAS41-/- cell lines with two disrupted GAS41 alleles can be generated following substitution of the endogenous gene by stable integration of GAS41 cDNA controlled by a tetracycline-regulated CMV promoter. Inactivation of this promoter by tetracycline withdrawal results in rapid depletion of GAS41, causing a significant decrease in RNA synthesis and subsequently cell death. Thus, our results indicate that GAS41 is required for RNA transcription.
Zimmermann
Katrin
K
Institut für Tierzucht und Tierverhalten, Dörnbergstrasse 25-27, 29223 Celle, Germany.
Ahrens
Katja
K
Matthes
Siegfried
S
Buerstedde
Jean-Marie
JM
Strätling
Wolf H
WH
Phi-van
Loc
L
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002470:Cell Survival / Q000502:physiology*; D002645:Chickens; D017931:DNA Primers; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*; D014162:Transfection
D017931:DNA Primers; D014157:Transcription Factors; C109446:YEATS4 protein, human