Holistic gene expression and total RNA quantification after Anti-Sense Agonist DNA Application to RNA Strands (ASADARS) procedure in Hek293 cells

Hybridization of oligonucleotides is effectively used to manipulate transcription level of cells. Here we introduce the method of hybridizing complementary single strand DNA (cDNA) to the RNA strands. We call this namely, Anti-Sense Agonist DNA Application to RNA Strands (ASADARS) method. The purpose of our study is to determine the effects of hybridization of single strand DNA oligomer to RNA transcript, to show specific or holistic interference. It is similar to siRNA, but DNA oligomers are replaced for interference instead of small RNA molecules. Identical to the initial protocol in RT-PCR, Hek293 cells were lysed and their RNA transcripts and were separated to synthesize cDNA. The RNA was removed, to leave only complementary ‘single strand DNA` oligomers. These DNA oligomers were then transfected back to 293 cells. For Controls, SH-Sy5y cDNA, 293 RNA, SH-Sy5y RNA, reagent and blank controls were treated. After 24 hours, the total RNA was decreased but the overall average microarray gene expression was up-regulated in 1.4 fold, in 30,926 genes. The single strand cDNA treatment, in specific gene target condition or holistic miscellaneous gene target was both evaluated. On the oligonucleotide treatment, either RNA or DNA delayed proliferation. Total protein level was not significantly altered. With this approach, primarily, the stability issues of siRNAs can be resolved by replacing it to DNA, which shows long term DNA-RNA hybridization. Cellular homeostasis and RNA compensation response is further postulated as a response of prolonged gene expression inhibitory or regulatory effects.