5 end phosphorylation reaction  (New England Biolabs)

Journal: Scientific Reports

Article Title: A RNA nanotechnology platform for a simultaneous two-in-one siRNA delivery and its application in synergistic RNAi therapy

doi: 10.1038/srep32363

Figure Lengend Snippet: ( A ) Rational design of ssDNA template for co-delivery of MDR1 siRNA and BCL2 siRNA. ( B ) Schematic illustration demonstrating self-assembly of Dsi RNA nanoparticles (Dsi RNPs) via rolling circle transcription (RCT) and one-step hybridization with folate-DNA-cholesterol fragments (FA-DNA-CHOL). RNA transcripts polymerized by T7 RNA polymerase-mediated RCT reaction consist of multiple tandem repeats of RNA hairpins, which contain the alternate sequences of BCL2 and MDR1 siRNA. Through base pairing between RNA and DNA, FA-DNA-CHOL fragments are sparsely tethered to RNA transcripts, and consequently, the amphiphilic RNA transcripts/FA-DNA-CHOL complexes are able to self-assemble into the highly packed RNA nanoparticles, which are feasible for co-delivery of MDR1 and BCL2 siRNA to the same cancer cells.

Article Snippet: T7 promoter primer, amino-cholesteryl (5′- and 3′-)-conjugated DNA, 3′-Cy5 labeled DNA, monomeric siRNA, and primers for qRT-PCR were all obtained from Bioneer Co. (Korea), as well as 5′-phosphorylated ssDNA templates for RCT reaction of various RNA nanoparticles including Dsi RNPs.

Techniques: Hybridization