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Only 1.5% of the human genome codes for protein. More and more evidence shows that the other 98.5% of the so-called junk DNA actually codes for small or large non-coding RNAs that regulate cellular activities. In addition to the milestones of chemical and protein drugs, it is predicted that a third milestone in drug development is RNA as drugs or drugs targeting to RNA (Guo et al., Adv Drug Deliv Rev. 2014 66:74). Currently the yield and cost of RNA nanoparticle or RNA drug production has been a barricade for the advancement of the RNA field in both research and clinical translation due to the multiple tedious manufacturing steps. For example, with a 98.5% of incorporation efficiency of chemical synthesis of a 100-nucleotide, RNA oligos will result in 78% contamination of the aborted bi-products. Here we report the one-pot production of RNA nanoparticles via automated processing and self-assembly. The continuous production of RNA by rolling circle transcription (RCT) using circular dsDNA template is coupled with the self-cleaving ribozyme encoded in the concatemeric RNA transcripts. Production was monitored in real time. Automatic production of RNA fragments enabled their assembly either in situ or via one-pot co-transcription to obtain RNA nanoparticles of desired motifs and functionalities from bottom-up assembly of multiple RNA fragments. In combination with the RNA nanoparticle construction process, a purification method using large scale electrophoresis column was also developed.
This article was published in the following journal.
Name: ACS nano
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Places provided for the processing, fabrication, assembly, and production of a line of products, services, commodities, or merchandise.
Nanometer-sized particles that are nanoscale in three dimensions. They include nanocrystaline materials; NANOCAPSULES; METAL NANOPARTICLES; DENDRIMERS, and QUANTUM DOTS. The uses of nanoparticles include DRUG DELIVERY SYSTEMS and cancer targeting and imaging.
Nanoparticles produced from metals whose uses include biosensors, optics, and catalysts. In biomedical applications the particles frequently involve the noble metals, especially gold and silver.
A layer of protein coating adsorbed by NANOPARTICLES upon entry into PLASMA or other protein-containing biological fluids, which affects how nanoparticles are internalized by cells and cleared from the body.
RNA transcripts of the DNA that are in some unfinished stage of post-transcriptional processing (RNA PROCESSING, POST-TRANSCRIPTIONAL) required for function. RNA precursors may undergo several steps of RNA SPLICING during which the phosphodiester bonds at exon-intron boundaries are cleaved and the introns are excised. Consequently a new bond is formed between the ends of the exons. Resulting mature RNAs can then be used; for example, mature mRNA (RNA, MESSENGER) is used as a template for protein production.
Bioinformatics is the application of computer software and hardware to the management of biological data to create useful information. Computers are used to gather, store, analyze and integrate biological and genetic information which can then be applied...