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  1. Article ; Online: A tool for optimizing messenger RNA sequence.

    Blakney, Anna K

    Nature

    2023  Volume 621, Issue 7978, Page(s) 262–264

    MeSH term(s) RNA, Messenger/genetics ; Biotechnology
    Chemical Substances RNA, Messenger
    Language English
    Publishing date 2023-05-30
    Publishing country England
    Document type News
    ZDB-ID 120714-3
    ISSN 1476-4687 ; 0028-0836
    ISSN (online) 1476-4687
    ISSN 0028-0836
    DOI 10.1038/d41586-023-01745-z
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Voices in Molecular Pharmaceutics: Meet Dr. Anna Blakney, an Engineer Who Seeks To Develop the Next Generation of Gene Therapies.

    Blakney, Anna K

    Molecular pharmaceutics

    2022  Volume 19, Issue 5, Page(s) 1231–1232

    MeSH term(s) Biopharmaceutics ; Genetic Therapy
    Language English
    Publishing date 2022-04-15
    Publishing country United States
    Document type Editorial
    ZDB-ID 2138405-8
    ISSN 1543-8392 ; 1543-8384
    ISSN (online) 1543-8392
    ISSN 1543-8384
    DOI 10.1021/acs.molpharmaceut.2c00257
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Self-amplifying RNA COVID-19 vaccine.

    Wayne, Christopher J / Blakney, Anna K

    Cell

    2024  Volume 187, Issue 8, Page(s) 1822–1822.e1

    Abstract: In November 2023, Japan's Ministry of Health, Labour and Welfare granted regulatory approval of ARCT-154, a self-amplifying RNA COVID-19 vaccine developed by Arcturus Therapeutics. Clinical trials showed comparable safety and efficacy using a lower dose ... ...

    Abstract In November 2023, Japan's Ministry of Health, Labour and Welfare granted regulatory approval of ARCT-154, a self-amplifying RNA COVID-19 vaccine developed by Arcturus Therapeutics. Clinical trials showed comparable safety and efficacy using a lower dose compared to the mRNA vaccine BNT162b2. To view this Bench-to-Bedside, open or download the PDF.
    MeSH term(s) Humans ; BNT162 Vaccine ; COVID-19/prevention & control ; COVID-19 Vaccines ; mRNA Vaccines/administration & dosage ; mRNA Vaccines/adverse effects
    Chemical Substances BNT162 Vaccine ; COVID-19 Vaccines ; mRNA Vaccines ; ARCT-154 vaccine
    Language English
    Publishing date 2024-04-15
    Publishing country United States
    Document type Journal Article
    ZDB-ID 187009-9
    ISSN 1097-4172 ; 0092-8674
    ISSN (online) 1097-4172
    ISSN 0092-8674
    DOI 10.1016/j.cell.2024.03.018
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Trends in the synthetic polymer delivery of RNA.

    Friesen, Josh J / Blakney, Anna K

    The journal of gene medicine

    2024  Volume 26, Issue 2, Page(s) e3672

    Abstract: Ribonucleic acid (RNA) has emerged as one of the most promising therapeutic payloads in the field of gene therapy. There are many unique types of RNA that allow for a range of applications including vaccination, protein replacement therapy, autoimmune ... ...

    Abstract Ribonucleic acid (RNA) has emerged as one of the most promising therapeutic payloads in the field of gene therapy. There are many unique types of RNA that allow for a range of applications including vaccination, protein replacement therapy, autoimmune disease treatment, gene knockdown and gene editing. However, RNA triggers the host immune system, is vulnerable to degradation and has a low proclivity to enter cells spontaneously. Therefore, a delivery vehicle is required to facilitate the protection and uptake of RNA therapeutics into the desired host cells. Lipid nanoparticles have emerged as one of the only clinically approved vehicles for genetic payloads, including in the COVID-19 messenger RNA vaccines. While lipid nanoparticles have distinct advantages, they also have drawbacks, including strong immune stimulation, complex manufacturing and formulation heterogeneity. In contrast, synthetic polymers are a widely studied group of gene delivery vehicles and boast distinct advantages, including biocompatibility, tunability, inexpensiveness, simple formulation and ease of modification. Some classes of polymers enhance efficient transfection efficiency, and lead to lower stimulation of the host immune system, making them more viable candidates for non-vaccine-related applications of RNA medicines. This review aims to identify the most promising classes of synthetic polymers, summarize recent research aimed at moving them into the clinic and postulate the future steps required for unlocking their full potential.
    MeSH term(s) RNA ; Polymers ; Nanoparticles ; Gene Transfer Techniques ; Transfection ; Genetic Therapy
    Chemical Substances RNA (63231-63-0) ; Polymers
    Language English
    Publishing date 2024-02-21
    Publishing country England
    Document type Journal Article ; Review
    ZDB-ID 1458024-x
    ISSN 1521-2254 ; 1099-498X
    ISSN (online) 1521-2254
    ISSN 1099-498X
    DOI 10.1002/jgm.3672
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Immune response to the components of lipid nanoparticles for ribonucleic acid therapeutics.

    Chen, Sunny P / Blakney, Anna K

    Current opinion in biotechnology

    2023  Volume 85, Page(s) 103049

    Abstract: Ribonucleic acid therapeutics have advantages over biologics and small molecules, including lower safety risks, cheaper costs, and extensive targeting flexibility, which is rapidly fueling the expansion of the field. This is made possible by ... ...

    Abstract Ribonucleic acid therapeutics have advantages over biologics and small molecules, including lower safety risks, cheaper costs, and extensive targeting flexibility, which is rapidly fueling the expansion of the field. This is made possible by breakthroughs in the field of drug delivery, wherein lipid nanoparticles (LNPs) are one of the most clinically advanced systems. LNP formulations that are currently approved for clinical use typically contain an ionizable cationic lipid, a phospholipid, cholesterol, and a polyethylene glycol-lipid; each contributes to the stability and/or effectiveness of LNPs. In this review, we discuss the immunomodulatory effects associated with each of the lipid components. We highlight several studies in which the components of LNPs have been implicated in cellular sensing and explore the pathways involved.
    MeSH term(s) RNA ; Lipids ; Liposomes ; Nanoparticles ; Immunity ; RNA, Small Interfering/metabolism
    Chemical Substances Lipid Nanoparticles ; RNA (63231-63-0) ; Lipids ; Liposomes ; RNA, Small Interfering
    Language English
    Publishing date 2023-12-19
    Publishing country England
    Document type Journal Article ; Review
    ZDB-ID 1052045-4
    ISSN 1879-0429 ; 0958-1669
    ISSN (online) 1879-0429
    ISSN 0958-1669
    DOI 10.1016/j.copbio.2023.103049
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: DNA vaccines join the fight against COVID-19.

    Blakney, Anna K / Bekker, Linda-Gail

    Lancet (London, England)

    2022  Volume 399, Issue 10332, Page(s) 1281–1282

    MeSH term(s) COVID-19/prevention & control ; Humans ; Vaccines, DNA/therapeutic use
    Chemical Substances Vaccines, DNA
    Language English
    Publishing date 2022-03-29
    Publishing country England
    Document type Journal Article ; Comment
    ZDB-ID 3306-6
    ISSN 1474-547X ; 0023-7507 ; 0140-6736
    ISSN (online) 1474-547X
    ISSN 0023-7507 ; 0140-6736
    DOI 10.1016/S0140-6736(22)00524-4
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Effect of in vitro transcription conditions on yield of high quality messenger and self-amplifying RNA.

    Popova, Petya G / Lagace, Melissa A / Tang, George / Blakney, Anna K

    European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V

    2024  Volume 198, Page(s) 114247

    Abstract: Messenger RNA (mRNA) and self-amplifying RNA (saRNA) vaccines against SARS-CoV-2 produced using in vitro transcription (IVT) were clinically approved in 2020 and 2022, respectively. While the industrial production of mRNA using IVT has been extensively ... ...

    Abstract Messenger RNA (mRNA) and self-amplifying RNA (saRNA) vaccines against SARS-CoV-2 produced using in vitro transcription (IVT) were clinically approved in 2020 and 2022, respectively. While the industrial production of mRNA using IVT has been extensively optimized, the optimal conditions for saRNA have been explored to a lesser extent. Most T7 polymerase IVT protocols have been specifically optimized for mRNA which is ∼5-10-fold smaller than saRNA and may have profound effects on both the quality and yield of longer transcripts. Here, we optimized IVT conditions for simultaneously increasing the yield of full-length transcripts and reducing dsRNA formation through Design of Experiments. Using a definitive screening approach, we found that the key parameters are temperature and magnesium in the outcome of RNA quality (% full length transcript) and yield in small scale synthesis. The most important parameter for reducing dsRNA formation for both mRNA and saRNA was Mg
    MeSH term(s) Humans ; RNA ; COVID-19 Vaccines ; RNA, Double-Stranded ; RNA, Messenger/genetics
    Chemical Substances RNA (63231-63-0) ; COVID-19 Vaccines ; RNA, Double-Stranded ; RNA, Messenger
    Language English
    Publishing date 2024-03-09
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 1065368-5
    ISSN 1873-3441 ; 0939-6411
    ISSN (online) 1873-3441
    ISSN 0939-6411
    DOI 10.1016/j.ejpb.2024.114247
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article ; Online: A duplex droplet digital PCR assay for absolute quantification and characterization of long self-amplifying RNA.

    Casmil, Irafasha C / Huang, Cynthia / Blakney, Anna K

    Scientific reports

    2023  Volume 13, Issue 1, Page(s) 19050

    Abstract: Self-amplifying messenger ribonucleic acid (saRNA) provides extended expression of genes of interest by encoding an alphavirus-derived RNA replicase and thus is 2-3 times larger than conventional messenger RNA. However, quality assessment of long RNA ... ...

    Abstract Self-amplifying messenger ribonucleic acid (saRNA) provides extended expression of genes of interest by encoding an alphavirus-derived RNA replicase and thus is 2-3 times larger than conventional messenger RNA. However, quality assessment of long RNA transcripts is challenging using standard techniques. Here, we utilized a multiplex droplet digital polymerase chain reaction (ddPCR) assay to assess the quality of saRNA produced from an in vitro transcription reaction and the replication kinetics in human cell lines. Using the one-step reverse transcription ddPCR, we show that an in vitro transcription generates 50-60% full-length saRNA transcripts. However, we note that the two-step reverse transcription ddPCR assay results in a 20% decrease from results obtained using the one-step and confirmed using capillary gel electrophoresis. Additionally, we provided three formulas that differ in the level of stringency and assumptions made to calculate the fraction of intact saRNA. Using ddPCR, we also showed that subgenomic transcripts of saRNA were 19-to-108-fold higher than genomic transcripts at different hours post-transfection of mammalian cells in copies. Therefore, we demonstrate that multiplex ddPCR is well suited for quality assessment of long RNA and replication kinetics of saRNA based on absolute quantification.
    MeSH term(s) Animals ; Humans ; RNA ; Polymerase Chain Reaction/methods ; RNA, Messenger/genetics ; Cell Line ; Biological Assay ; Real-Time Polymerase Chain Reaction/methods ; Mammals
    Chemical Substances RNA (63231-63-0) ; RNA, Messenger
    Language English
    Publishing date 2023-11-03
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-023-46314-6
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Clinical perspective on topical vaccination strategies.

    G Popova, Petya / Chen, Sunny P / Liao, Suiyang / Sadarangani, Manish / Blakney, Anna K

    Advanced drug delivery reviews

    2024  Volume 208, Page(s) 115292

    Abstract: Vaccination is one of the most successful measures in modern medicine to combat diseases, especially infectious diseases, and saves millions of lives every year. Vaccine design and development remains critical and involves many aspects, including the ... ...

    Abstract Vaccination is one of the most successful measures in modern medicine to combat diseases, especially infectious diseases, and saves millions of lives every year. Vaccine design and development remains critical and involves many aspects, including the choice of platform, antigen, adjuvant, and route of administration. Topical vaccination, defined herein as the introduction of a vaccine to any of the three layers of the human skin, has attracted interest in recent years as an alternative vaccination approach to the conventional intramuscular administration because of its potential to be needle-free and induce a superior immune response against pathogens. In this review, we describe recent progress in developing topical vaccines, highlight progress in the development of delivery technologies for topical vaccines, discuss potential factors that might impact the topical vaccine efficacy, and provide an overview of the current clinical landscape of topical vaccines.
    MeSH term(s) Humans ; Vaccination ; Vaccines ; Skin ; Adjuvants, Immunologic ; Antigens
    Chemical Substances Vaccines ; Adjuvants, Immunologic ; Antigens
    Language English
    Publishing date 2024-03-24
    Publishing country Netherlands
    Document type Journal Article ; Review
    ZDB-ID 639113-8
    ISSN 1872-8294 ; 0169-409X
    ISSN (online) 1872-8294
    ISSN 0169-409X
    DOI 10.1016/j.addr.2024.115292
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: A duplex droplet digital PCR assay for absolute quantification and characterization of long self-amplifying RNA

    Irafasha C. Casmil / Cynthia Huang / Anna K. Blakney

    Scientific Reports, Vol 13, Iss 1, Pp 1-

    2023  Volume 12

    Abstract: Abstract Self-amplifying messenger ribonucleic acid (saRNA) provides extended expression of genes of interest by encoding an alphavirus-derived RNA replicase and thus is 2–3 times larger than conventional messenger RNA. However, quality assessment of ... ...

    Abstract Abstract Self-amplifying messenger ribonucleic acid (saRNA) provides extended expression of genes of interest by encoding an alphavirus-derived RNA replicase and thus is 2–3 times larger than conventional messenger RNA. However, quality assessment of long RNA transcripts is challenging using standard techniques. Here, we utilized a multiplex droplet digital polymerase chain reaction (ddPCR) assay to assess the quality of saRNA produced from an in vitro transcription reaction and the replication kinetics in human cell lines. Using the one-step reverse transcription ddPCR, we show that an in vitro transcription generates 50–60% full-length saRNA transcripts. However, we note that the two-step reverse transcription ddPCR assay results in a 20% decrease from results obtained using the one-step and confirmed using capillary gel electrophoresis. Additionally, we provided three formulas that differ in the level of stringency and assumptions made to calculate the fraction of intact saRNA. Using ddPCR, we also showed that subgenomic transcripts of saRNA were 19-to-108-fold higher than genomic transcripts at different hours post-transfection of mammalian cells in copies. Therefore, we demonstrate that multiplex ddPCR is well suited for quality assessment of long RNA and replication kinetics of saRNA based on absolute quantification.
    Keywords Medicine ; R ; Science ; Q
    Subject code 612
    Language English
    Publishing date 2023-11-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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