Assembly of multifunctional phi29 pRNA nanoparticles for specific delivery of siRNA and other therapeutics to targeted cells

Haque

et al. 2013

RNA

Self Cite

142

237

Due to structural flexibility, RNase sensitivity, and serum instability, RNA nanoparticles with concrete shapes for in vivo application remain challenging to construct. Here we report the construction of 14 RNA nanoparticles with solid shapes for targeting cancers specifically. These RNA nanoparticles were resistant to RNase degradation, stable in serum for >36 h, and stable in vivo after systemic injection. By applying RNA nanotechnology and exemplifying with these 14 RNA nanoparticles, we have established the technology and developed "toolkits" utilizing a variety of principles to construct RNA architectures with diverse shapes and angles. The structure elements of phi29 motor pRNA were utilized for fabrication of dimers, twins, trimers, triplets, tetramers, quadruplets, pentamers, hexamers, heptamers, and other higher-order oligomers, as well as branched diverse architectures via hand-in-hand, foot-to-foot, and arm-on-arm interactions. These novel RNA nanostructures harbor resourceful functionalities for numerous applications in nanotechnology and medicine. It was found that all incorporated functional modules, such as siRNA, ribozymes, aptamers, and other functionalities, folded correctly and functioned independently within the nanoparticles. The incorporation of all functionalities was achieved prior, but not subsequent, to the assembly of the RNA nanoparticles, thus ensuring the production of homogeneous therapeutic nanoparticles. More importantly, upon systemic injection, these RNA nanoparticles targeted cancer exclusively in vivo without accumulation in normal organs and tissues. These findings open a new territory for cancer targeting and treatment. The versatility and diversity in structure and function derived from one biological RNA molecule implies immense potential concealed within the RNA nanotechnology field.

Section: Resultsmentioning

confidence: 99%

Fabrication of 14 different RNA nanoparticles for specific tumor targeting without accumulation in normal organs

Haque

et al. 2013

RNA

Self Cite

142

237

DNA and RNA Nanotechnology

“…In particular, computationally designed [72] and experimentally tested [32,35,73,74] nanorings take advantage of the so-called RNA kissing loop ineracting motifs (Figure 1, case 4). Other examples use the structural motifs extracted from the phage phi29 pRNAs [75] that were widely used in the engineering of multiple stable and functional RNA nanoscaffolds [76][77][78][79][80][81][82][83][84][85][86][87]. An alternative to the tecto-RNA designing strategy is exemplified by the nanocubes (Figure 2) [88,89].…”

Section: Multifunctional Rna and Dna Nanoparticlesmentioning

confidence: 99%

Triggering RNAi with multifunctional RNA nanoparticles and their delivery

Dao¹,

Viard²,

Martins³

et al. 2015

in the bloodstream, poor cellular uptake, and inefficient intracellular release. In an attempt to solve these issues, different types of RNAi therapeutic delivery strategies including multifunctional RNA nanoparticles are being developed. In this mini-review, we will briefly describe some of the current approaches.Keywords: RNA nanotechnology, RNA nanoparticles, RNA/DNA hybrids, RNA interference, siRNA, delivery IntroductionAccording to the Social Security Administration, average Americans that reach age 65 today most likely will live to be 84 years old [1]. However, with older age, the chances of contracting deadly diseases, such as cancer, increases dramatically. Some cancers (e.g. breast cancer) can be removed surgically but this does not guarantee that the disease will not return within a patient's lifetime. For other types of cancer (e.g. chronic lymphocytic leukemia), surgery may have very little effect (http://www. cancer.org). Other available treatments are chemo-and immunotherapies. However, these alternatives lack target specificity and cause severe toxic side effects affecting the growth of hair, nails, loss of appetite and blood cell count, just to name a few (http://www.cancer.org). Therefore, the advancements in biomedical technologies that provide safe and effective cancer treatment are in demand. Among the novel approaches is the recognition and use of specific intracellular RNA signatures (e.g. an overexpression of certain genes) that are especially important in detection and personalized treatments of cancers as well as viral infections, and autoimmune diseases [2][3][4]. The wide use of novel therapeutics based on target specific RNA-mediated gene silencing, called RNA interference or RNAi, will likely become the next breakthrough in cancer therapy. The first successful therapeutic knockdown of the endogenous gene, apolipoprotein B (ApoB), occurred in a 2004 study [5], only a few years after the original discovery of RNAi [6]. In 2010, DOI 10.1515/rnan-2015-0001 Received April 6, 2015 accepted May 6, 2015 Abstract: Proteins are considered to be the key players in structure, function, and metabolic regulation of our bodies. The mechanisms used in conventional therapies often rely on inhibition of proteins with small molecules, but another promising method to treat disease is by targeting the corresponding mRNAs. In 1998, Craig Mellow and Andrew Fire discovered dsRNA-mediated gene silencing via RNA interference or RNAi. This discovery introduced almost unlimited possibilities for new gene silencing methods, thus opening new doors to clinical medicine. RNAi is a biological process that inhibits gene expression by targeting the mRNA. RNAi-based therapeutics have several potential advantages (i) a priori ability to target any gene, (ii) relatively simple design process, (iii) sitespecificity, (iv) potency, and (v) a potentially safe and selective knockdown of the targeted cells. However, the problem lies within the formulation and delivery of RNAi therapeutics including rapid excretion, instab...

DNA and RNA Nanotechnology

“…The second class encompasses tectonic units that can be split into structural motifs and interacting motifs possessing discrete secondary and tertiary structures which can be combined to rationally produce self-assembling nanoparticles with predictable, multi-dimensional structures in vivo or in vitro. Some examples of structural motifs are double-stranded RNA helices, C-loops [72] (which increase the helical twists), kink-turns [97], right angle motifs [78], tRNA motifs [98] (the latter three bends within an oligonucleotide strand); as well as three [76,99], four [100,101], and five way junctions [100]. Examples of interacting motifs are tetraloopreceptors [74] (an interaction between a four-base hairpin "tetra-loop" motif and a "receptor" structured internal loop motif), kissing loops [102,103] (coaxial stacking motifs), sticky ends [104,105], and paranemic motifs [81] (an interaction of "crosshatched" stacking helices).…”

Section: Rna Architectonic Approach (Tectorna)mentioning

confidence: 99%

“…It has greatly enhanced simplicity and biocompatibility when compared with peptide, liposomal, or synthetic nanomaterial conjugation for RNA-based functionalities; RNA nanoparticles can be equipped with custom tailored combinations of therapeutic agents or biosensors, which show promise to improve targeting and penetration of cells or tissues safely and efficiently [76,95,107], provide "cocktail"-like multi-target therapies [106], advanced imaging or imaging concurrent with therapeutics [110]. In addition, they can do so while guaranteeing strict control of the 3D orientation and stoichiometry of those agents [102,104].…”

Section: Rna Architectonic Approach (Tectorna)mentioning

confidence: 99%

“…The breadth of functionality in biologically occurring RNAs provides a very powerful foundation for the bottom up design of nanodevices in the form of various structural motifs and functional elements which operate in native biological systems, and can be derived from X-ray and NMR structures of natural molecules [59,[71][72][73][74][75][76][77][78]. This repertoire of natural RNA motifs is augmented by new motifs with novel binding selectivities or enzymatic capabilities, obtained by in vitro selection, also called Systematic Evolution of Ligands by Exponential Enrichment (SELEX) [79,80], in which randomized pools of oligonucleiotides are iteratively selected, amplified, and mutated to enrich highaffinity binding to targets; as well as through rational design from biochemical principles [74,77,81].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Engineered RNA Nanodesigns for Applications in RNA Nanotechnology

Afonin¹,

Lindsay²,

Shapiro³

2013

IntroductionThe physical world presents a very different landscape at the nanometer scale. Physical phenomena such as inertia and gravity vanish from view; instead the interactions of matter and energy are dominated by other phenomena like wave-particle duality, uncertainty, and quantum electrodynamics. Nanotechnology is the engineering of functional systems at this scale, where the axioms that govern material and device design differ dramatically from those found at the macroscale level. More precisely, "nanotechnology" typically encompasses both the miniaturization of existing technologies to the nanoscale, and the discipline of engineering molecular-scale systems from the bottom up, producing constructs with fundamentally new qualities that revolutionize human engineering, from materials and manufacturing, electronics, and information technology, to medicine, biotechnology, energy, and even security. The array of goals and applications in nanotechnology intersects with an equally wide array of techniques and materials with their own emergent properties in the field. One such branch is concerned with the re-engineering of biological mechanisms, systems, and molecules in new forms with new utilities, broadly termed bio-nanotechnology.The field of nanotechnology taking advantage of nucleic acids has its origins in the work of Nadrian Seeman and coworkers who have, over the past 30 years, spearheaded the development of DNA nano-object fabrication utilizing DNA self-assembly [1][2][3][4][5][6]. Briefly, DNA nanotechnology uses the nature of DNA complementarity for the construction of DNA tiles with discrete secondary structure by canonical WatsonCrick interactions (G-C and A-T base pairing), using a relatively small number of structural rules fundamentally based on Holliday junction motifs. The use of these rules has resulted in the engineering and characterization of numerous DNA 3D nanoscaffolds with different connectiviities [7][8][9][10][11][12][13][14][15][16][17][18] and the ability for some of them to promote targeted delivery by functioning as DNA nano-capsules [19][20][21] or DNA nano-carriers for other functionialities [22]. Another powerful technique called DNA "origami", developed by Paul Rothemund [23], has been extended from its original scope for designing different 2D DNA shapes [24] and functional templates [25][26][27][28] Abstract Nucleic acids have emerged as an extremely promising platform for nanotechnological applications because of their unique biochemical properties and functions. RNA, in particular, is characterized by relatively high thermal stability, diverse structural flexibility, and its capacity to perform a variety of functions in nature. These properties make RNA a valuable platform for bio-nanotechnology, specifically RNA Nanotechnology, that can create de novo nanostructures with unique functionalities through the design, integration, and re-engineering of powerful mechanisms based on a variety of existing RNA structures and their fundamental biochemical properties. This review...