TransIT¢ç-Oligo Transfection ReagentA high efficiency, low toxicity, transfection reagent optimized for oligonucleotide delivery into a wide range of cell types
Figure 1. The TransIT¢ç-Oligo Reagent Achieves High Transfection Efficiency. HeLa cells transfected using TransIT¢ç-Oligo Reagent and Label IT¢ç Cy¢ç3 and Label IT¢ç Fluorescein labeled phosphothioate DNA oligos in complete media for 24 hours.
Figure 2. The TransIT¢ç-Oligo Reagent Effectively Transfects a 2'OMe RNA Oligo that Blocks a Cryptic Splice Site. The HeLa-Luc 705 reporter cell line (Kang et al. 1998, 37:6235) used in this study contains a luciferase reporter construct that has the ©¬-globin 705 intron inserted into the luciferase ORF. A mutation present at position 705 of the ©¬-globin intron activates two cryptic splice sites within the intron that lead to the production of a spliced luciferase mRNA that is disrupted by a small intron with an in-frame stop codon, thus preventing translation of functional luciferase protein. The transfection of a 2'OMe oligonucleotide (TriLink BioTechnologies, Inc.) complementary to the cryptic 705 splice site inhibits splicing at the cryptic splice sites enabling the complete removal of the ©¬-globin intron and production of a mRNA with a complete, uninterrupted luciferase ORF.
The HeLa-Luc 705 cell line was transfected with increasing amounts of the anti-705 splice site 2'OMe RNA oligo at the indicated final concentrations using the TransIT¢ç-Oligo Transfection Reagent. The cells were harvested 24 hours post-transfection and assayed for luciferase activity. The increase in luciferase activity indicates effective delivery of the anti-705 splice site RNA oligo using the TransIT¢ç-Oligo Reagent.
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