scholarly journals Anionic Complex with Efficient Expression and Good Safety Profile for mRNA Delivery

Pharmaceutics ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 126
Author(s):  
Eri Hamada ◽  
Tomoaki Kurosaki ◽  
Junya Hashizume ◽  
Hitomi Harasawa ◽  
Hiroo Nakagawa ◽  
...  
Keyword(s):  
Protein Expression ◽  
Messenger Rna ◽  
Charge Ratio ◽  
Attractive Alternative ◽  
Target Cells ◽  
Anionic Complex ◽  
In Vitro Protein Expression ◽  
Vitro Protein

We previously found that a complex comprising plasmid DNA (pDNA), polyethylenimine (PEI), and γ-polyglutamic acid (γ-PGA) had high transgene efficiency without cytotoxicity in vitro and in vivo. However, messenger RNA (mRNA) remains an attractive alternative to pDNA. In this study, we developed a safe and effective delivery system for mRNA to prevent its degradation and efficiently deliver it into target cells. Various cationic and anionic complexes were produced containing PEI, γ-PGA, and an mRNA encoding firefly luciferase. Their physicochemical properties and cytotoxicities were analyzed and the in vitro and in vivo protein expression were determined. The cationic mRNA/PEI complex showed high in vitro protein expression with strong cytotoxicity. The anionic complex was constructed as mRNA/PEI8/γ-PGA12 complex with a theoretical charge ratio of 1:8:12 based on the phosphate groups of the mRNA, nitrogen groups of PEI, and carboxylate groups of γ-PGA. It was stable and showed high in vitro protein expression without cytotoxicity. After intravenous administration of mRNA/PEI8/γ-PGA12 complex to mice, high protein expression was observed in the spleen and liver and slight expression was observed in the lung over 24 h. Thus, the newly constructed mRNA/PEI8/γ-PGA12 complex provides a safe and effective strategy for the delivery of mRNA.

An Overview of nano delivery systems for targeting RNA interference-based therapy in ulcerative colitis

2021 ◽  
Vol 27 ◽  
Author(s):  
Iman Alfagih ◽  
Basmah Aldosari ◽  
Bushra AlQuadeib ◽  
Alanood Almurshedi ◽  
Murtaza Tambuwala
Keyword(s):  
Ulcerative Colitis ◽  
Rna Interference ◽  
Protein Expression ◽  
Messenger Rna ◽  
Delivery Systems ◽  
Target Cells ◽  
Intracellular Space ◽  
Therapeutic Benefits ◽  
Negative Effect

: Ulcerative colitis (UC) is one of the main subtypes of inflammatory bowel disease. UC has a negative effect on patients’ quality of life, and it is an important risk factor for the development of colitis-associated cancer. Patients with UC need to take medications for their entire life because no permanent cure is available. Therefore, approaches that target messenger RNA (mRNA) of proinflammatory cytokines or anti-inflammatory cytokines are needed to improve the safety of UC therapy and promote intestinal mucosa recovery. The major challenge facing RNA interference-based therapy is the delivery of RNA molecules to the intracellular space of target cells. Moreover, nonspecific and systemic protein expression inhibition can result in adverse effects and less therapeutic benefits. Thus, it is important to develop an efficient delivery strategy targeting the cytoplasm of target cells to avoid side effects caused by off-target protein expression inhibition. This review focuses on the most recent advances in the targeted nano delivery systems of siRNAs and mRNA that have shown in vivo efficacy.

A PMMA microfluidic droplet platform for in vitro protein expression using crude E. coli S30 extract

Lab on a Chip ◽  
2009 ◽  
Vol 9 (23) ◽  
pp. 3391 ◽  
Author(s):  
N. Wu ◽  
Y. Zhu ◽  
S. Brown ◽  
J. Oakeshott ◽  
T. S. Peat ◽  
...  
Keyword(s):  
Protein Expression ◽  
E Coli ◽  
S30 Extract ◽  
In Vitro Protein Expression ◽  
Vitro Protein

Proficient Target Selection in Structural Genomics by In Vitro Protein Expression on Gateway Recombination Plasmids

2002 ◽  
pp. 197-202 ◽  
Author(s):  
Vincent Monchois ◽  
Renaud Vincentelli ◽  
Céline Deregnaucourt ◽  
Chantal Abergel ◽  
Jean-Michel Claverie
Keyword(s):  
Protein Expression ◽  
Structural Genomics ◽  
Target Selection ◽  
In Vitro Protein Expression ◽  
Vitro Protein

scholarly journals High-Yield, In Vitro Protein Expression Using a Continuous-Exchange, Coupled Transcription/Translation System

BioTechniques ◽  
2001 ◽  
Vol 31 (4) ◽  
pp. 948-953 ◽  
Author(s):  
G.A. Martin ◽  
R. Kawaguchi ◽  
Y. Lam ◽  
A. DeGiovanni ◽  
M. Fukushima ◽  
...  
Keyword(s):  
Protein Expression ◽  
High Yield ◽  
Translation System ◽  
In Vitro Protein Expression ◽  
Vitro Protein

scholarly journals A nucleotidyltransferase toxin inhibits growth of Mycobacterium tuberculosis through inactivation of tRNA acceptor stems

2020 ◽  
Vol 6 (31) ◽  
pp. eabb6651 ◽  
Author(s):  
Yiming Cai ◽  
Ben Usher ◽  
Claude Gutierrez ◽  
Anastasia Tolcan ◽  
Moise Mansour ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Bacterial Toxins ◽  
Human Pathogen ◽  
Biochemical Activity ◽  
Unknown Mechanism ◽  
In Vitro Protein Expression ◽  
Block Protein Synthesis ◽  
Vitro Protein

Toxin-antitoxin systems are widespread stress-responsive elements, many of whose functions remain largely unknown. Here, we characterize the four DUF1814-family nucleotidyltransferase-like toxins (MenT1–4) encoded by the human pathogen Mycobacterium tuberculosis. Toxin MenT3 inhibited growth of M. tuberculosis when not antagonized by its cognate antitoxin, MenA3. We solved the structures of toxins MenT3 and MenT4 to 1.6 and 1.2 Å resolution, respectively, and identified the biochemical activity and target of MenT3. MenT3 blocked in vitro protein expression and prevented tRNA charging in vivo. MenT3 added pyrimidines (C or U) to the 3′-CCA acceptor stems of uncharged tRNAs and exhibited strong substrate specificity in vitro, preferentially targeting tRNASer from among the 45 M. tuberculosis tRNAs. Our study identifies a previously unknown mechanism that expands the range of enzymatic activities used by bacterial toxins, uncovering a new way to block protein synthesis and potentially treat tuberculosis and other infections.

scholarly journals Application of In Vitro Protein Expression to Human Prote

2002 ◽  
Vol 2 ◽  
pp. 73-74 ◽  
Author(s):  
Peter T. Beernink ◽  
Simone S. Krupka ◽  
Victoria Lao ◽  
George Martin ◽  
Matthew A. Coleman
Keyword(s):  
Protein Expression ◽  
In Vitro Protein Expression ◽  
Vitro Protein
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