Related Experiment Video

Updated: Jul 16, 2025

In Vitro Synthesis of Modified mRNA for Induction of Protein Expression in Human Cells
10:07

In Vitro Synthesis of Modified mRNA for Induction of Protein Expression in Human Cells

Published on: November 13, 2014

25.0K

Evaluation of self-amplifying mRNA platform for protein expression and genetic stability: Implication for mRNA

Swarda Deo1, Kaushik Desai1, Aishwarya Patare1

  • 1Gennova Biopharmaceuticals Ltd. ITBT Park, Hinjawadi Phase 2 Road, Hinjawadi Rajiv Gandhi Infotech Park, Hinjawadi, Pune, Maharashtra, 411057, India.

Biochemical and Biophysical Research Communications
|September 22, 2023
PubMed

Insights

Self-amplifying mRNA (SAM) platforms show sustained antigenic protein expression without genomic integration or mutations. This research supports the potential of alphavirus-based SAM technology for future applications.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Vaccinology

Background:

  • The widespread use of messenger RNA (mRNA) vaccines has raised questions regarding the long-term expression, safety, and potential for genomic integration of mRNA vaccine technologies.
  • Concerns exist about the stability and potential unintended effects of mRNA platforms in vaccine development.

Purpose of the Study:

  • To evaluate the duration of antigenic protein expression from self-amplifying mRNA (SAM) constructs (mRNA-614 and mRNA-S1LC).
  • To investigate the potential for genomic integration and mutations in SAM-based mRNA platforms.
  • To explore the structural and functional aspects of non-structural proteins (nsPs) within the SAM system.

Main Methods:

  • Utilized Expi-293F™, HEK-293T, and ARPE-19 cell lines for mRNA expression studies.
  • Assessed protein expression using sandwich-ELISA, flow cytometry (FACS), luciferase activity assays, and Western blotting.
  • Confirmed mRNA integrity through sequencing and assessed genomic integration via PCR analysis.
  • Generated in silico 3D structures of nsPs and expressed these proteins using the T7 system.

Main Results:

  • Sustained antigenic protein expression was observed from the mRNA-614 and mRNA-S1LC SAM constructs.
  • Sequencing confirmed that the transcribed mRNA remained un-mutated throughout the study.
  • No evidence of reverse transcribed mRNA integration into the host cell genome was detected up to 7 days post-transfection.
  • High-quality 3D structures of nsPs were successfully generated and their genes cloned and expressed.

Conclusions:

  • The alphavirus-based SAM platform demonstrates stable and long-lasting antigenic protein expression.
  • The absence of genomic integration and mutations supports the safety profile of this SAM platform.
  • These findings highlight the significant potential of alphavirus-based SAM technology as a future vaccine modality.
Keywords:
And genome integrationNon-structural proteins (nsPs)Self-amplifying mRNAmRNA vaccine

More Related Videos

A Nonviral Approach to Generate Transient Chimeric Antigen Receptor T Cells Using mRNA for Cancer Immunotherapy09:56

A Nonviral Approach to Generate Transient Chimeric Antigen Receptor T Cells Using mRNA for Cancer Immunotherapy

Published on: February 21, 2025

599
Efficient Transcriptionally Controlled Plasmid Expression System for Investigation of the Stability of mRNA Transcripts in Primary Alveolar Epithelial Cells10:49

Efficient Transcriptionally Controlled Plasmid Expression System for Investigation of the Stability of mRNA Transcripts in Primary Alveolar Epithelial Cells

Published on: March 6, 2020

6.0K

Related Concept Videos

mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

2.9K
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
942
What is Gene Expression?01:36

What is Gene Expression?

A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised  of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then...
8.6K
Regulation of Expression Occurs at Multiple Steps02:24

Regulation of Expression Occurs at Multiple Steps

Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
22.8K
RNA Stability01:53

RNA Stability

Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
33.6K
Microorganisms in Medicine and Therapeutics01:29

Microorganisms in Medicine and Therapeutics

Microorganisms play a fundamental role in vaccine development, gene therapy, and therapeutic production. Their biological properties are harnessed to advance medicine and public health. Beyond immunization, microorganisms contribute to gut health, antibiotic synthesis, and genetic disease treatment.Live Attenuated and Inactivated VaccinesLive attenuated vaccines, such as the measles, mumps, and rubella (MMR) vaccine, utilize weakened forms of pathogens to closely resemble natural infections.
49