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A Reverse Transcription Nucleic-Acid-Based Barcoding System for In Vivo Measurement of Lipid Nanoparticle mRNA
Kevin C Wang1, Tiana L Young1, Jingan Chen2
1Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario M5S 3M2, Canada.
ACS Bio & Med Chem Au
|February 24, 2025
Summary
We developed Quantitative Analysis of Reverse Transcribed Barcodes (QuART), a new method to screen lipid nanoparticle (LNP) delivery systems for mRNA therapeutics. This nucleic acid-based system enables efficient in vitro and in vivo functional LNP screening.
Area of Science:
- Biotechnology
- Molecular Biology
- Drug Delivery Systems
Background:
- Lipid nanoparticles (LNPs) are crucial for mRNA therapeutics delivery, as seen in COVID-19 vaccines.
- Current LNP screening methods hinder the rapid development of novel mRNA therapeutics.
- Efficient and scalable screening is essential for advancing LNP technology.
Purpose of the Study:
- To develop a novel nucleic acid-based system for assessing functional mRNA delivery by lipid nanoparticles (LNPs).
- To enable high-throughput screening of LNP formulations for improved mRNA therapeutic development.
Main Methods:
- Developed Quantitative Analysis of Reverse Transcribed Barcodes (QuART), a system linking mRNA delivery to a cDNA barcode readout.
- Utilized a bacterial retron reverse transcription system for functional mRNA delivery measurement.
- Validated QuART for assessing LNP delivery efficiency in vitro (cell culture) and in vivo (mice).
Main Results:
- QuART successfully measured functional mRNA delivery in both cell culture and live mice.
- The system demonstrated its capability to identify effective LNP delivery.
- Demonstrated the potential for multiplexing QuART for enhanced screening capacity.
Conclusions:
- QuART provides a novel and effective method for evaluating LNP-mediated mRNA delivery.
- This system can accelerate the discovery and optimization of LNP formulations for mRNA therapeutics.
- QuART facilitates high-throughput screening, addressing a key bottleneck in LNP development.
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