TransIT®-Oligo Transfection Reagent

TransIT®-Oligo Reagent Achieves High Transfection Efficiency. HeLa cells transfected using TransIT®-Oligo Reagent and Label IT® Cy®3 and Label IT® Fluorescein labeled phosphothioate DNA oligos in complete media for 24 hours.

TransIT®-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 ß-globin 705 intron inserted into the luciferase ORF. A mutation present at position 705 of the ß-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 ß-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®-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®-Oligo Reagent.