Biotinylated photocleavable polyethylenimine: capture and triggered release of nucleic acids from solid supports.
A biotinylated photocleavable polyethylenimine (B-PC-PEI) was designed and synthesized for the capture and controlled release of nucleic acids from solid supports. B-PC-PEI was synthesized via a three-step reaction process and verified by 1H NMR and mass spectrometry. In aqueous solution, the o-nitrobenzyl group within B-PC-PEI was efficiently cleaved by 5 min of 365 nm light exposure from a distance of 20 cm (9 mW/cm2). When coupled to streptavidin-coated beads, the PEI domain of Cy5-labeled B-PC-PEI was released by 365 nm light exposure. In contrast, a Cy5-labeled biotinylated PEI (B-PEI) was used as a control and negligible fluorescence loss was observed. Cy5-labeled siRNA was electrostatically captured to streptavidin-coated beads preabsorbed with B-PC-PEI or B-PEI, and flow cytometry demonstrated significant loss of fluorescence from the bead surface after 5 min of light exposure only for B-PC-PEI, demonstrating controlled release of siRNA from the bead surface. Finally, the release of the Cy5-labeled siRNA into the supernatant was quantified. The release of Cy5-siRNA into the supernatant was significantly greater after 5 min of light exposure for B-PC-PEI/streptavidin beads compared to 0 min exposure and remained unchanged for B-PEI/streptavidin beads. B-PC-PEI facilitates capture and triggered release of surface-tethered nucleic acids with light exposure and is fully compatible with streptavidin-based applications.
Handwerger
Rachel G
RG
Institute for Medicine and Engineering, 1024 Vagelos Laboratories, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Diamond
Scott L
SL
D001710:Biotin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D019921:Biotinylation; D002232:Carbocyanines / Q000737:chemistry; D018014:Gene Transfer Techniques; D008863:Microspheres; D010782:Photolysis; D011073:Polyamines / Q000138:chemical synthesis / Q000737:chemistry*; D011094:Polyethyleneimine / Q000737:chemistry; D034741:RNA, Small Interfering / Q000737:chemistry*; D019809:Streptavidin / Q000737:chemistry
D002232:Carbocyanines; D011073:Polyamines; D034741:RNA, Small Interfering; C085321:cyanine dye 5; D001710:Biotin; D011094:Polyethyleneimine; D019809:Streptavidin