WO2023038433A1 - Arn circulaire et son utilisation - Google Patents

Arn circulaire et son utilisation Download PDF

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Publication number
WO2023038433A1
WO2023038433A1 PCT/KR2022/013454 KR2022013454W WO2023038433A1 WO 2023038433 A1 WO2023038433 A1 WO 2023038433A1 KR 2022013454 W KR2022013454 W KR 2022013454W WO 2023038433 A1 WO2023038433 A1 WO 2023038433A1
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vector
circular rna
site
internal ribosome
ribosome entry
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Korean (ko)
Inventor
박영근
조병문
카무투마니
조영란
허훈
김신영
송민아
김보영
정지윤
이지원
조현준
정미선
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Geneone Life Science Inc
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Geneone Life Science Inc
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression

Definitions

  • the present invention relates to circular RNA and its use, and more particularly to circular RNA that is more stable than linear RNA in vivo.
  • circRNA circular RNA
  • ecircRNA exonic RNA
  • EIcircRNA exon-intron RNA
  • ciRNA intronic RNA
  • ecircRNA is the major circRNA and is mainly produced in vivo by a process called backsplicing.
  • EcircRNAs are mainly located in the cytoplasm and have a variety of functions.
  • the best-known function of circRNAs is as a microRNA (miRNA) sponge, such as the circRNA CDR1as and Sry.
  • circRNAs have also been demonstrated to serve as protein sponges and scaffolds for protein complexes.
  • circRNAs also play a role in regulating the translation and stability of mRNA levels and the activity of proteins.
  • circRNAs as biomarkers have been demonstrated to be associated with age-dependent neuronal accumulation in Drosophila, and are known to be involved in cancer and other diseases.
  • the present inventors have completed the present invention by developing a circular RNA platform with improved stability compared to the existing platform.
  • an object of the present invention is a promoter; internal ribosomal entry site (IRES); protein coding region; 3' untranslated region (UTR); poly A; And to provide a vector for production of a circular RNA comprising a 5' group I intron fragment including a 5' splicing site.
  • Another object of the present invention is an internal ribosome entry site; protein coding region; 3' UTR; poly A; and a 5' group I intronic fragment including a 5' splicing site.
  • Another object of the present invention is a vector for producing the circular RNA; Or to provide a pharmaceutical composition for the prevention or treatment of a target disease comprising; or the circular RNA.
  • Another object of the present invention is a vector for producing the circular RNA; Or to provide a vaccine composition for preventing a target disease comprising; or the circular RNA.
  • Another object of the present invention is to provide a method for expressing a protein of interest in a cell, including the step of transducing the vector or the circular RNA for production of the circular RNA into the cell.
  • Another object of the present invention is to provide a method for treating a target disease comprising administering the vector for producing the circular RNA or the circular RNA to a subject in need thereof.
  • the present invention provides a vector for the production of circular RNA comprising the following elements operably linked:
  • the present invention also provides a circular RNA comprising the following elements operably linked:
  • the present invention is a vector for producing the circular RNA; Or the circular RNA; It provides a pharmaceutical composition for the prevention or treatment of a target disease, including.
  • the present invention is a vector for producing the circular RNA; Or the circular RNA; It provides a vaccine composition for preventing a target disease comprising.
  • the present invention provides a method for expressing a target protein in a cell, comprising transducing a vector for production of the circular RNA or the circular RNA into the cell.
  • the present invention is a vector for producing the circular RNA; Or, administering the circular RNA to a subject in need thereof; provides a method for treating a disease of interest, including the.
  • the circular RNA according to the present invention has higher stability than linear RNA even after treatment with RNase and higher stability than linear RNA even after long-term storage. This means that since the circular RNA of the present invention has high stability in vitro or in vivo, it can maintain the expression of the target protein for a long time in vitro or in vivo. Therefore, the circular RNA of the present invention is a platform for preventing or treating a target disease; Or a vaccine platform for preventing infection of a target disease; it can be used in various ways.
  • FIG. 1 is a diagram showing the results of confirming mRNA-S, which is a linear RNA, and circRNA-S, which is a circular RNA, through electrophoresis.
  • Figure 2 is a diagram showing the results of confirming the stability of mRNA-S, which is a linear RNA, and circRNA-S, which is a circular RNA, according to RNase treatment.
  • FIG. 3 is a diagram showing the results of confirming the stability of mRNA-S, which is a linear RNA, and circRNA-S, which is a circular RNA, over time.
  • Figure 4 is a diagram showing the results of confirming the splicing site of pCircRNA without a 3' splicing site through splicing junction site sequencing.
  • the invention provides a vector for the production of circular RNA comprising the following elements operably linked:
  • operatively linked means a functional linkage between a nucleotide expression control sequence (eg, a promoter sequence) and another nucleotide sequence. Accordingly, the regulatory sequence may thereby regulate the transcription and/or translation of the other nucleotide sequence.
  • a nucleotide expression control sequence eg, a promoter sequence
  • a vector means a means for expressing a target gene in a host cell.
  • viral vectors such as plasmid vectors, cosmid vectors and bacteriophage vectors, adenoviral vectors, retroviral vectors and adeno-associated viral vectors.
  • the usable vectors include plasmids often used in the art (e.g., pGLS, pSC101, pGV1106, pACYC177, ColE1, pKT230, ME290, pBR322, pUC8/9, pUC6, pBD9, pHC79, pIJ61, pLAFR1, pHV14, pGEX series, pET series and pUC19, etc.), phages (eg, ⁇ gt4 ⁇ B, ⁇ Charon, ⁇ z1, and M13, etc.) or viruses (eg, CMV, SV40, etc.).
  • plasmids often used in the art (e.g., pGLS, pSC101, pGV1106, pACYC177, ColE1, pKT230, ME290, pBR322, pUC8/9, pUC6, pBD9, pHC79, pIJ61, pLAFR1, pHV14,
  • the vector of the present invention can typically be constructed as a vector for cloning or a vector for expression.
  • the vector those conventionally used in the art to express foreign proteins in plants, animals, or microorganisms may be used.
  • the vector may be constructed through various methods known in the art.
  • the vector may be constructed using a prokaryotic or eukaryotic cell as a host.
  • the origins of replication operating in the eukaryotic cell included in the vector are the f1 origin of replication, the SV40 origin of replication, the pMB1 origin of replication, the adeno origin of replication, the AAV origin of replication, the CMV origin of replication, and the BBV Including the origin of replication, etc., but is not limited thereto.
  • promoters derived from the genome of mammalian cells eg, metallotionine promoter
  • promoters derived from mammalian viruses eg, adenovirus late promoter, vaccinia virus 7.5K promoter, SV40 promoter,
  • the cytomegalovirus (CMV) promoter and the TK promoter of HSV can be used, and usually have a polyadenylation sequence as a transcription termination sequence.
  • a protein coding region means a gene encoding a target protein.
  • the protein coding region may be a region encoding a protein having therapeutic efficacy, an antigen protein, an antibody, and the like, and any gene encoding a target protein may be applied.
  • the vector preferably does not contain a 3' splicing site upstream of the internal ribosome entry site.
  • the structure of a preferred vector of the present invention is shown as pCircRNA-ver2.0 in FIG. 4 .
  • the vector may be one in which the elements 1) to 6) are sequentially operably linked.
  • the vector comprises the following elements operably linked:
  • 3'UTR represented by the nucleotide sequence of SEQ ID NO: 3;
  • variants of the above-described nucleotide sequences are included within the scope of the present invention.
  • the variant of the base sequence has a sequence homology of at least 70%, more preferably at least 80%, even more preferably at least 90%, and most preferably at least 95% with the base sequence described in the sequence listing.
  • It means a sequence that exhibits substantially the same physiological activity as the nucleotide sequence described in the sequence listing.
  • the "percentage of sequence homology" for polynucleotides is determined by comparing two optimally aligned sequences with a comparison region, wherein a portion of the polynucleotide sequence in the comparison region is a reference sequence (addition or deletion) for the optimal alignment of the two sequences. may include additions or deletions (i.e., gaps) compared to (not including).
  • the 5' group I intronic fragment containing the 5' splicing site is preferably derived from Anabaena pre-tRNA.
  • the internal ribosome entry site may be derived from Encephalomyocarditis virus (EMCV), and any internal ribosome entry site exhibiting the same activity can be applied without limitation.
  • EMCV Encephalomyocarditis virus
  • the poly A is preferably 50 to 300 bp, more preferably 100 to 200 bp, and most preferably 151 bp.
  • the vector preferably includes a modified nucleic acid, natural nucleic acid or non-natural nucleic acid, but is not limited thereto.
  • the vector can produce circular RNA by self-splicing, and it is preferable that the 5' end of the internal ribosome entry site is cleaved by self-splicing, and further Preferably, 1 to 50 nt of the 5' end of the internal ribosome entry site may be cut by self-splicing, and more preferably, 28 nt may be cut.
  • the internal ribosome entry site of the vector preferably overlaps the 5' end of the internal ribosome entry site with the 5' splicing site, more preferably the 5' end of the internal ribosome entry site. It may overlap with the 5' splicing site by 1 to 10 nt, most preferably by 7 nt.
  • the present invention provides a circular RNA comprising the following elements operably linked:
  • the circular RNA is preferably a circular RNA comprising the following elements operably linked:
  • 3'UTR represented by the nucleotide sequence of SEQ ID NO: 3;
  • the circular RNA is preferably cleaved from 1 to 50 nt of the 5' end of the internal ribosome entry site, more preferably 28 nt from the 5' end.
  • the circular RNA preferably has the 5' end of the internal ribosome entry site overlapping the 5' splicing site, more preferably the circular RNA has the 5' end of the internal ribosome entry site It may overlap 1 to 10 nt with the 5' splicing site, and most preferably may overlap 7 nt.
  • the circular RNA according to the present invention has much higher stability than linear RNA under RNase treatment conditions and long-term storage conditions. This means that the circular RNA of the present invention can stably and continuously express the target protein under various conditions. Therefore, the circular RNA of the present invention may be used in medicine for preventing or treating a target disease; Vaccine use for preventing infection of target disease; And expression method of the target protein; can be used as a platform technology in various fields including.
  • the present invention provides a vector for the production of circular RNA; Or circular RNA; It provides a pharmaceutical composition for the prevention or treatment of a target disease comprising.
  • Vectors and circular RNAs for the production of circular RNAs of the present invention include protein coding regions.
  • the protein coding region refers to a gene encoding a target protein, and when the target protein exhibits a preventive or therapeutic effect of a specific disease (ie, target disease), it can be usefully used for medicinal purposes for the target disease.
  • the present invention provides a vector for producing the circular RNA; Or the circular RNA; It provides a vaccine composition for preventing a target disease comprising.
  • a vector for producing the circular RNA; and circular RNA; the protein coding region included in may be a gene encoding an antigen protein.
  • the gene encoding the antigen protein may be a spike gene represented by the nucleotide sequence of SEQ ID NO: 5 used in Examples of the present invention, and the scope of the present invention is not limited thereto.
  • the vaccine is a veterinary vaccine containing an antigenic material, and is administered for the purpose of inducing active or passive immunity specific to a target disease.
  • the "immunologically effective amount" of PCV2 means an amount sufficient to exhibit a preventive effect of related diseases and an amount sufficient to not cause side effects or severe or excessive immune reactions, and the exact dosage concentration varies depending on the specific immunogen to be administered. It can be easily determined by a person skilled in the art according to factors well-known in the medical field, such as age, weight, health, sex, sensitivity to drugs of the subject, administration route, and administration method of the vaccination subject, and can be administered once or several times. can
  • the vaccine composition according to the present invention includes a vector for producing circular RNA as an active ingredient; and circular RNA; In addition, one or more immune enhancers or excipients or carriers suitable for constituting the vaccine composition may be included.
  • An adjuvant that may be included in the vaccine composition of the present invention refers to a substance that enhances the immune response of an injected animal, and many different adjuvants are known to those skilled in the art.
  • the immune enhancers include Freund's complete and incomplete immune enhancers, vitamin E, nonionic blocking polymers, muramyl dipeptide, Quil A, mineral oil and non-mineral oil and Carbopol, water-in-oil emulsion immune enhancers, etc., but are limited thereto it is not going to be
  • Carriers that may be included in the vaccine composition of the present invention are known to those skilled in the art, and include, but are not limited to, proteins, sugars, and the like.
  • the above carriers may be aqueous or non-aqueous solutions, suspensions, and emulsions.
  • non-aqueous carriers include propylene glycol, polyethylene glycol, edible oils such as olive oil, and injectable organic esters such as ethyl oleate.
  • Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media.
  • Parenteral carriers include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's or fixed oils.
  • Carriers for intravenous injection include electrolyte replenishers, liquid and nutritional supplements, and the like, such as those based on Ringer's dextrose.
  • the vaccine composition of the present invention may further contain preservatives and other additives such as, for example, antimicrobial agents, antioxidants, chelating agents, inert gases, and the like.
  • the preservatives include formalin, thimerosal, neomycin, polymyxin B and amphotericin B, and the like.
  • the vaccine composition of the present invention may include one or more suitable emulsifiers, such as Span or Tween.
  • the vaccine composition of the present invention may include a protecting agent, and a protecting agent known in the art may be used without limitation, which may include lactose (LPGG) or trehalose (TPGG), It is not limited thereto.
  • the present invention provides a method for expressing a target protein in a cell, including transducing a vector for production of the circular RNA or the circular RNA into a cell.
  • the vector and circular RNA for production of the circular RNA of the present invention include a protein coding region, and genes encoding various target proteins can be applied to the protein coding region.
  • Circular RNA according to the present invention has much higher stability than linear RNA under RNase treatment conditions and long-term storage conditions, which means that the circular RNA of the present invention can stably and continuously express a target protein in cells.
  • a vector for producing the circular RNA or a method for treating a target disease comprising administering the circular RNA to a subject in need thereof is provided.
  • the subject is a subject expected to develop a disease in which the target protein exhibits therapeutic efficacy; diseased individuals; Alternatively, it may be an object that has been cured, but is not limited thereto.
  • the target protein refers to a protein encoded by a 'protein coding region' included in a vector or circular RNA for circular RNA production.
  • Redundant content is omitted in consideration of the complexity of the present specification, and terms not otherwise defined in the present specification have meanings commonly used in the technical field to which the present invention belongs.
  • pCircDNA-S and pCircDNA-eGFP were used as vectors for circular RNA production, and circular RNAs (CircDNA-S and CircDNA-eGFP) were produced using these vectors.
  • a vector for producing circular RNA includes 1) a promoter represented by the nucleotide sequence of SEQ ID NO: 1; 2) an internal ribosome entry site represented by the nucleotide sequence of SEQ ID NO: 2; 3) protein coding regions; 4) 3'UTR represented by the nucleotide sequence of SEQ ID NO: 3; 5) Poly A; and 6) a 5' Group I intron fragment including a 5' splicing site represented by the nucleotide sequence of SEQ ID NO: 4; characterized in that they are sequentially operably linked.
  • the pCircDNA-S of this experiment applied the spike gene represented by the nucleotide sequence of SEQ ID NO: 5 to the protein coding region, and the eGFP gene represented by the nucleotide sequence of SEQ ID NO: 7 was applied to pCircDNA-eGFP.
  • the 5' group I intron fragment including the 5' splicing site is derived from Anabaena pre-tRNA, and the internal ribosome entry site is derived from Encephalomyocarditis virus (EMCV).
  • poly A included in the vector is 151 bp.
  • linear RNA (mRNA-S, mRNA-eGFP) produced using vectors pDNA-S and pDNA-eGFP for linear RNA production was used.
  • the vectors pDNA-S and pDNA-eGFP for the production of the linear RNA are represented by the nucleotide sequences of SEQ ID NOs: 15 and 17, respectively, and the linear RNAs mRNA-S and mRNA-eGFP produced using them are SEQ ID NOs: 15 and 17, respectively. is represented by the nucleotide sequence of
  • IVT is the process of synthesizing RNA using T7 RNA polymerase in vitro using linearized template DNA. IVT was performed using linearized DNA according to the manufacturer's instructions of the EZTM MEGA T7 Transcription Kit (Enzynomics). Since pDNA-S and pDNA-eGFP are linear RNA forms, ARCA (8 mM) (New England Biolabs) was mixed and reacted in IVT process for mRNA stability. Each sample was reacted at 37 ° C for 1 hour and 40 minutes, and DNase I (2 ⁇ l / 1 rxn) was added and reacted at 37 ° C for 20 minutes.
  • RNA was purified using the Lithium Chloride (LiCl) Precipitation Solution included in the EZTM MEGA T7 Transcription Kit (Enzynomics). LiCl Precipitation Solution was added as much as half the volume of the reaction solution, thoroughly mixed, and maintained at -20 ° C for more than 30 minutes. It was then centrifuged at 4° C. for 15 minutes at the highest possible speed, and the supernatant was carefully removed. For washing, 700 ⁇ l of 70% EtOH was added and centrifuged again under the same conditions as above. After completely removing EtOH, the RNA pellet was resuspended in RNase-free water.
  • LiCl Precipitation Solution was added as much as half the volume of the reaction solution, thoroughly mixed, and maintained at -20 ° C for more than 30 minutes. It was then centrifuged at 4° C. for 15 minutes at the highest possible speed, and the supernatant was carefully removed. For washing, 700 ⁇ l of 70% EtOH was added and centrifuged again
  • the circular RNAs circRNA-S and circRNA-eGFP are characterized in that 28 nt of the 5' end of the internal ribosome entry site is cleaved and the 5' end of the internal ribosome entry site overlaps the 5' splicing site by 7 nt. to be
  • RNA electrophoresis was performed in the same manner as in Example 1-3. As a control group in this experiment, mRNA-S and CircRNA R -S were used. The circRNA R -S is RNA in circular form synthesized in a compact form from Houston Cincinnati Leading Medicine. RNA electrophoresis results are shown in FIG. 2 .
  • HEK293T cells were cultured in 10% FBS (Gibco), 1% penicillin/streptomycin, and Dulbecco's Modified Eagle Medium (DMEM) high glucose medium at 37°C and 5% CO 2 . 1 ⁇ g each of mRNA-eGFP or circRNA-eGFP was added to HEK293T cells according to the manufacturer's instructions. After that, Lipofectamine2000 (Invitrogen) and Opti-MEM (Gibco) were used to transfect 500,000 cells/1mL per well in a 6-well plate. After 4 hours of transfection, the medium was changed to the first culture medium, DMEM high glucose medium.
  • FBS Gibco
  • DMEM Dulbecco's Modified Eagle Medium
  • the RNAs extracted from the lysed cells are mRNA-eGFP (SEQ ID NO: 17) in linear RNA form and circRNA-eGFP (SEQ ID NO: 13) in circular RNA form.
  • RNA 1 ⁇ g was used to synthesize cDNA according to the manufacturer's instructions of the RevertAid First Strand cDNA Synthesis Kit (Thermo Scientific).
  • Realtime qPCR was performed using the synthesized cDNA to confirm the gene expression level according to the target gene.
  • Faststart SYBR green master (Roche)
  • 10X concentrated solution of the PCR primer was prepared.
  • Forward and reverse primers were used at a concentration of 0.4 ⁇ M
  • eGFP was used as the target gene
  • b-actin was used as the housekeeping gene.
  • qPCR was performed using a Lightcycler96 device (Roche). The Ct value for each sample was compared and calculated, and the gene expression level was normalized to b-actin, a housekeeping gene. The qPCR results are shown in FIG. 3 .
  • splicing junction sequencing 10 ⁇ L of splicing reaction column-purified circRNA with RNase R, 1 ⁇ L of oligodT (invitrogen), 1 ⁇ L of dNTP (Enzynomics), and 1 ⁇ L of nuclease free water (13 ⁇ L final volume) were added to 65 The secondary structure was normalized by heating at °C for 5 min and cooling on ice for 3 min.
  • the reverse transcription reaction was performed according to the manufacturer's instructions using 1 ⁇ L of SuperiorScript III reverse transcriptase (Enzynomics) for the internal region of the putative circRNA, and the specific conditions are shown in Table 3 below.
  • PCR products for sequencing were synthesized using 2X topsimple Dyemix - Tenuto (Enzynomics) and a pair of primers spanning splice junctions. Specific PCR conditions are shown in Table 4.
  • Step Temperature(°C) Time One 98 3:00 2 98 0:10 3 64 0:30 touch down(-0.5°C) 4 72 0:30 5 GOTO STEP2 9X 6 98 0:10 7 59 0:30 8 72 0:30 9 GOTO STEP 14X 10 72 2:00 11 4 ⁇
  • the synthesized DNA was purified according to the manufacturer's instructions of a DNA cleanup kit (New England Biolabs). Sanger-sequencing was requested to the company to confirm the splicing junction site, and the results are shown in FIG. 4 .

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Abstract

La présente invention concerne un ARN circulaire et son utilisation, et plus particulièrement un ARN circulaire plus stable in vivo qu'un ARN linéaire. Des expériences ont confirmé que l'ARN circulaire, selon la présente invention, possède une plus grande stabilité qu'un ARN linéaire, même lorsqu'il est traité à la RNase, et qu'il possède une plus grande stabilité qu'un ARN linéaire, même lorsqu'il est stocké pendant une longue période. Ceci signifie que l'ARN circulaire de la présente invention présente une stabilité in vitro ou in vivo élevée et permet ainsi de maintenir l'expression in vitro ou in vivo d'une protéine cible pendant une longue durée. Ainsi, l'ARN circulaire de la présente invention peut être utilisé de diverses manières en tant que plate-forme pour la prévention ou le traitement d'une maladie cible, ou en tant que plate-forme vaccinale pour la prévention d'une infection d'une maladie cible.
PCT/KR2022/013454 2021-09-07 2022-09-07 Arn circulaire et son utilisation Ceased WO2023038433A1 (fr)

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CN116694630B (zh) * 2023-08-03 2023-10-31 呈诺再生医学科技(北京)有限公司 一种促进环状rna过表达的序列组合及其应用

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Publication number Priority date Publication date Assignee Title
WO2014186334A1 (fr) * 2013-05-15 2014-11-20 Robert Kruse Traduction intracellulaire d'arn circulaire
WO2017222911A1 (fr) * 2016-06-20 2017-12-28 The Board Of Trustees Of The Leland Stanford Junior University Arn circulaires et leur utilisation dans l'immunomodulation
WO2019094486A1 (fr) * 2017-11-07 2019-05-16 The University Of North Carolina At Chapel Hill Méthodes et compositions pour des molécules d'arn circulaire
KR20210018323A (ko) * 2018-06-06 2021-02-17 매사추세츠 인스티튜트 오브 테크놀로지 진핵 세포에서의 번역을 위한 원형 rna
KR20210057019A (ko) * 2018-07-24 2021-05-20 메이오 파운데이션 포 메디칼 에쥬케이션 앤드 리써치 원형화된 조작된 rna 및 방법

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014186334A1 (fr) * 2013-05-15 2014-11-20 Robert Kruse Traduction intracellulaire d'arn circulaire
WO2017222911A1 (fr) * 2016-06-20 2017-12-28 The Board Of Trustees Of The Leland Stanford Junior University Arn circulaires et leur utilisation dans l'immunomodulation
WO2019094486A1 (fr) * 2017-11-07 2019-05-16 The University Of North Carolina At Chapel Hill Méthodes et compositions pour des molécules d'arn circulaire
KR20210018323A (ko) * 2018-06-06 2021-02-17 매사추세츠 인스티튜트 오브 테크놀로지 진핵 세포에서의 번역을 위한 원형 rna
KR20210057019A (ko) * 2018-07-24 2021-05-20 메이오 파운데이션 포 메디칼 에쥬케이션 앤드 리써치 원형화된 조작된 rna 및 방법

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