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Current advances in Phi29 pRNA biology and its application in drug delivery

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Abstract Bacteriophage 29 (Phi29) packaging RNA (pRNA) is one of the key components in the viral DNA‐packaging motor. It contains two functional domains facilitating the translocation of DNA into the viral capsid by interacting with other elements in the motor and promoting adenosine triphosphates hydrolysis. Through the connection between interlocking loops in adjacent pRNA monomers, pRNA functions in the form of multimer ring in the motor. Previous studies have addressed the unique structure and conformation of pRNA. However, there are different DNA‐packaging models proposed for the viral genome transportation mechanism. The DNA‐packaging ability and the unique features of pRNA have been attracting efforts to study its potential applications in nanotechnology. The pRNA has been proved to be a promising tool for delivering nucleic acid‐based therapeutic molecules by covalent linkage with ribozymes, small interfering RNAs, aptamers, and artificial microRNAs. The flexibility in constructing dimers, trimers, and hexamers enables the assembly of polyvalent nanoparticles to carry drug molecules for therapeutic purposes, cell ligands for target delivery, image detector for drug entry monitoring, and endosome disrupter for drug release. Besides these fascinating pharmacological advantages, pRNA‐based drug delivery has also been demonstrated to prolong the drug half life with minimal induction of immune response and toxicity. WIREs RNA 2012, 3:469–481. doi: 10.1002/wrna.1111 This article is categorized under: RNA Structure and Dynamics > RNA Structure, Dynamics, and Chemistry RNA Structure and Dynamics > Influence of RNA Structure in Biological Systems RNA Processing > RNA Editing and Modification RNA in Disease and Development > RNA in Disease

Structure of the Phi29 DNA‐packaging system. The Phi29 genome packaging motor is consist of three components, pRNA, gp16, and gp3. The packaging motor interacts with the procapsid through the connector protein. The pRNA functions in a multimer complex (5‐mer or 6‐mer).

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Schematic structures of pRNA–ribozyme, pRNA–siRNA, pRNA–aptamers, and pRNA–AmiRs. (a) pRNA–ribozyme—the helical regions of pRNA and the ribozyme were connected by linkers. (b) pRNA–siRNA—the helical region of pRNA is replaced by a double‐stranded siRNA sequence. (c) pRNA–aptamers—the helical regions of pRNA and the aptamer are connected by linkers. (d) pRNA–AmiR—the helical region of pRNA is replaced by an AmiR sequences, which contains a partially complementary double strands.

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Secondary structures of pRNA monomers, dimers, and hexamers. (a) Sequence and secondary structure of pRNA. Two major functional domains are indicated in light green (DNA translocation domain) and light purple (procapsid‐binding domain). In the DNA translocation domain, the essential bulge for pRNA function is circled out. In the procapsid‐binding domain, three loops (R, L, and head loops) are indicated by white area. In R and L loops, the 4 nts involved in the pRNA loop–loop interaction are indicated in red (R loop) and brown (L loop). (b) Schematic structure of A/a′‐A/a′ pRNA dimer. Two identical pRNA A/a′ monomers interact each other through the base paring of sites on the R and L loops. (c) Schematic structure of A/a′ pRNA hexamers. Six pRNA A/a′ monomers interact each other through their loops to form a hexamer. (d) Schematic structure of A/b′‐B/a′ pRNA dimer. pRNA A/b′ and pRNA B/a′ monomers interact each other through the base paring of sites on the R and L loops.

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RNA Structure and Dynamics > RNA Structure, Dynamics, and Chemistry
RNA Structure and Dynamics > Influence of RNA Structure in Biological Systems
RNA in Disease and Development > RNA in Disease
RNA Processing > RNA Editing and Modification

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