DNazyme-mediated cleavage of Twist transcripts and increase in cellular apoptosis.
DNazymes is a group of catalytic nucleic acids that can be designed to cleave target mRNA molecules in a base-specific way. Twist is a basic helix-loop-helix transcription factor that is involved in the regulation of cellular differentiation and apoptosis. Moreover, it was shown to function in skull development and cause craniosynostosis. DZ-TWT DNazyme was designed to down-regulate Twist expression. The ability of DZ-TWT to cleave mouse Twist mRNA was first shown in a cell-free environment against full-length Twist mRNA. Following transfections of the DZ-TWT in C3H10T1/2 cells, a significant reduction of Twist mRNA levels was observed. This was accompanied by a significant rise in p21 mRNA levels. Finally, DZ-TWT transfections resulted in an increase of cellular apoptosis, demonstrating the importance of Twist in apoptotic pathways. These results prove the usefulness of DNazymes to characterize Twist gene function and further experiments in animals should demonstrate its complete physiological role.
Hjiantoniou
Eleni
E
The Cyprus Institute of Neurology and Genetics, P.O. Box 23462, 1683 Nicosia, Cyprus.
Iseki
Sachiko
S
Uney
James B
JB
Phylactou
Leonidas A
LA
D000818:Animals; D017209:Apoptosis / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D002460:Cell Line; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000235:genetics; D021881:DNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation; D066298:In Vitro Techniques; D051379:Mice; D009687:Nuclear Proteins / Q000235:genetics* / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors; D014158:Transcription, Genetic; D014162:Transfection; D051879:Twist-Related Protein 1; D015854:Up-Regulation
C493957:Cdkn1a protein, mouse; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D021881:DNA, Catalytic; D009687:Nuclear Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D051879:Twist-Related Protein 1