Japan Science and Technology Agency

Kyoto University

Tan Inoue

Hirohide Saito

Kei Endo

Kenichi Yoshikawa

Ayako Yamada

Yusho Kato

Toru Yamanaka

Shunichi Kashida

Karin Hayashi

Tetsuhiro Kobayashi

Tomoaki Hara

Hara; Tomoaki

A translational control method using an RNA-protein interaction motif is provided. The method comprises a step of introducing an mRNA having: a 5'UTR regulation structure comprising: (1) a cap structure at the 5&#x2032; terminus, (2) a spacer positioned on the 3&#x2032; side of the cap structure, and (3) one or more RNA motifs positioned on the 3&#x2032; side of the spacer, which comprises an RNA-protein interaction motif-derived nucleotide sequence or a variant thereof; and a nucleotide sequence encoding a target protein gene on the 3&#x2032; side of the 5&#x2032;UTR regulation structure, into a cell in the presence of a protein specifically binding to the RNA motifs, wherein a translational level is decreased as the number of bases of the spacer decreases, and the translational level is decreased as the number of the RNA motifs increases.

Translational control system using RNA-protein interaction motif

An object of the present invention is to provide a translationally regulatable mRNA which has wider application and can perform specific ON-OFF regulation, an RNA-protein complex specifically bound to the mRNA, and a translational regulatory system. The present invention provides an mRNA having an RNA-protein complex interacting motif-derived nucleotide sequence 5' to the ribosome-binding site or within the 5&#x2032; region of the open reading frame, and an mRNA having a nucleotide sequence complementary to an RNA-protein complex interacting motif-derived nucleotide sequence 5&#x2032; to the ribosome-binding site or within the 5&#x2032; region of the open reading frame.

Protein-responsive translational regulatory system using RNA-protein interacting motif

An object of the present invention is to construct an mRNA which specifically responds to a short RNA sequence and can activate, repress, and regulate the translation of the desired gene, and to construct an artificial cell model system using a liposome comprising the mRNA and a cell-free translational system encapsulated therein. The present invention provides: an mRNA comprising a target RNA-binding site located immediately 5' to the ribosome-binding site, and a nucleotide sequence located 5&#x2032; to the target RNA-binding site, the nucleotide sequence being complementary to the ribosome-binding site; an mRNA comprising a small RNA-binding site located 3&#x2032; to the start codon, and a nucleotide sequence located 3&#x2032; to the small RNA-binding site, the nucleotide sequence encoding a protein; and a liposome comprising any of these mRNAs encapsulated therein.

Small RNA-dependent translational regulatory system in cell or artificial cell model

An object of the present invention is to provide an RNAi control system using an RNA-protein interaction motif. The present invention provides an shRNA comprising: a guide strand having a sequence complementary to a target sequence; a passenger strand which forms a duplex with the guide strand; and a linker strand which links the guide strand and the passenger strand, wherein the linker strand comprises an RNP-derived protein-binding motif sequence. The present invention also provides an RNAi control system comprising: the shRNA; and an RNP-derived protein which specifically binds to a protein-binding motif sequence in the shRNA.

Construction of protein-responsive shRNA/RNAi control system using RNP motif

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Kyoto, Japan

Tan Inoue