Unknown,Transcriptomics,Genomics,Proteomics

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Gene profile in H1299 cells treated with PTD-DRBD GAPDH siRNA or treated with Lipofection GAPDH siRNA


ABSTRACT: Whole genome microarrays were probed with total mRNA from PTD-DRBD GAPDH siRNA treated H1299 cells at 12 h and 24 h. Using a 1.6x fold increase/decrease filter of cellular mRNAs, we detected a dramatic reduction in the target GAPDH mRNA along with a limited number of both up and down regulated genes. The up regulated genes were reduced in numbers and to nearly background 1.6x levels at 24 h, while the down regulated genes increased slightly in numbers and maintained a similar magnitude at 24 h. In contrast, lipofection treated cells showed both a dramatic increase in both the total number of genes altered and the magnitude of the increase. In addition, the numbers of genes affected increased between 12 h and 24 h, suggesting that lipofection of siRNAs into cells results in a substantial alteration to the transcriptome and may thereby confound interpretation of experimental outcomes. Moreover, the GAPDH specific knockdown was significantly smaller than PTD-DRBD mediated knockdown. Total RNA obtained from H1299 cells treated with PTD-DRBD GAPDH siRNA, Lipofection GAPDH siRNA or PBS (Control) after 12 or 24 hours post-treatment

ORGANISM(S): Homo sapiens

SUBMITTER: Steven Dowdy 

PROVIDER: E-GEOD-15723 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Efficient siRNA delivery into primary cells by a peptide transduction domain-dsRNA binding domain fusion protein.

Eguchi Akiko A   Meade Bryan R BR   Chang Yung-Chi YC   Fredrickson Craig T CT   Willert Karl K   Puri Nitin N   Dowdy Steven F SF  

Nature biotechnology 20090517 6


RNA interference (RNAi) induced by short interfering RNA (siRNA) allows for discovery research and large-scale screening; however, owing to their size and anionic charge, siRNAs do not readily enter cells. Current approaches do not deliver siRNAs into a high percentage of primary cells without cytotoxicity. Here we report an efficient siRNA delivery approach that uses a peptide transduction domain-double-stranded RNA-binding domain (PTD-DRBD) fusion protein. DRBDs bind to siRNAs with high avidit  ...[more]

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