RNA Delivery via DNA-Inspired Janus Base Nanotubes for Extracellular Matrix Penetration

Abstract

RNA delivery into deep tissues with dense extracellular matrix (ECM) has been challenging. For example, cartilage is a major barrier for RNA and drug delivery due to its avascular structure, low cell density and strong negative surface charge. Cartilage ECM is comprised of collagens, proteoglycans, and various other noncollagneous proteins with a spacing of 20nm. Conventional nanoparticles are usually spherical with a diameter larger than 50-60nm (after cargo loading). Therefore, they presented limited success for RNA delivery into cartilage. Here, we developed Janus base nanotubes (JBNTs, self-assembled nanotubes inspired from DNA base pairs) to assemble with small RNAs to form nano-rod delivery vehicles (termed as “Nanopieces”). Nanopieces have a diameter of ∼20nm (smallest delivery vehicles after cargo loading) and a length of ∼100nm. They present a novel breakthrough in ECM penetration due to the reduced size and adjustable characteristics to encourage ECM and intracellular penetration.

Comparison between Janus-Base Nanotubes and Carbon Nanotubes: A Review on Synthesis, Physicochemical Properties, and Applications

“…However, the structure is no longer called “K1”, as the side chain does not necessarily need to be lysine to be considered JBNt. The base units of JBNt are still the DNA-inspired base pairs, with success in using both guanine and cytosine (G^C) [ 2 ] or thymine and adenine (A^T) [ 23 ].…”

“…Zhou et al synthesized the JBNt monomer according to the previously published patent [ 2 ]. The synthesis of the monomer alone involves the intermediate steps A1–A9, with the final step using A9 to yield crude JBNt monomers, where it is subsequently purified by high-performance liquid chromatography (HPLC).…”

“…Among the current advances in engineering materials, nanotechnology has posed as a promising area of study for the purpose of enhanced drug and gene delivery, among other various applications. Nanoparticles come in a variety of morphologies and chemical compositions but are often spherical and have a diameter of ~50 nm or larger, limiting their ability for cargo delivery to cells or the extracellular matrix (ECM) [ 1 , 2 ]. As an alternative to spherical nanoparticles, nanotubes have generated a lot of interest over the past two decades, with carbon nanotubes (CNTs) showing some of the most potential due to their distinct and exceptional properties and wide array of applicability.…”

“…Yu and Chens to be more applicable to biomedical purposes [ 6 ]. In addition to cargo loading, the JBNt can be functionalized to enable targeted delivery [ 2 ], similar to CNTs. JBNts also have the ability to form a type of noncovalent linked nanoparticle called nanopieces (JBNps), which are multiple JBNt strands bonded together to form larger delivery vehicles, unlike CNT.…”

Nanomaterials for Protein Delivery in Anticancer Applications

“…For drug delivery purposes, RNTs or JBNTs can be processed into non-covalent nanoparticles (such as nanopieces or NPs). The NPs can be used to inhibit disease mRNA or miRNA expression [ 50 , 51 ]. In an anticancer application [ 51 ], NPs were shown to deliver oligonucleotides intracellularly to human tumor cells in vitro and in vivo and can inhibit expression of oncogenic miRNA, which in turn can restore the expression of RGS16.…”