Custom Mitochondrial Targeting mRNA Synthesis Service

Introduction

Mitochondria are vital organelles, often referred to as the "powerhouses of the cell," critical for energy production, metabolism, and cell survival. Dysfunctional mitochondria are implicated in a wide range of diseases, including neurodegenerative disorders, metabolic syndromes, and cancer. Precisely delivering therapeutic or research molecules to these organelles has historically been a significant challenge.

Creative Biolabs' Custom Mitochondrial Targeting mRNA Synthesis accelerates research and drug discovery via advanced in vitro transcription and precise targeting sequences, overcoming challenges in mitochondrial delivery and protein expression optimization.

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Custom Mitochondrial Targeting mRNA Synthesis

Mitochondrial Targeting mRNA (Mito-mRNA) represents a cutting-edge approach in gene therapy and research, designed to overcome the challenges of delivering functional proteins to mitochondria. By fusing a specific Mitochondrial Targeting Sequence (MTS) to the coding region of a gene of interest, the resulting mRNA molecule gains the ability to be selectively transported to and translated at the mitochondrial surface. This ensures that the newly synthesized protein is directly imported into the mitochondria, allowing for precise functional restoration or modulation. A commonly utilized and highly effective MTS is the N-terminal 23 amino acids of cytochrome oxidase subunit 8 precursor protein (MSVLTPLLLRGLTGSARRLPVPR), which efficiently guides the mRNA-protein complex to its mitochondrial destination.

Fig.1 Mitochondrial DNA (mtDNA) core proteins and peripheral proteins, along with other factors, are jointly involved in the organization, stability, and communication of mtDNA.¹

Workflow

Required Starting Materials: To initiate your project, we typically require:

• Target Gene Sequence: The full coding sequence of the protein you wish to express.
• Desired Mitochondrial Targeting Sequence (MTS): While we can provide standard MTS options (e.g., N-terminal 23 amino acids of cytochrome oxidase subunit 8 precursor protein), any specific or novel MTS you wish to incorporate should be provided.
• Specific Application Details: Information regarding your experimental model (e.g., cell lines, in vivo models) and the intended application (e.g., functional restoration, disease modeling, therapeutic development).

1. Synthesis

   We use a robust IVT platform: starting with a linearized DNA template, optimized reactions with quality reagents/enzymes yield full-length, high-fidelity mRNA. Controlled conditions enable modified nucleotide incorporation to boost stability, translation efficiency, and reduce immunogenicity.

2. Purification

   Creative Biolabs uses advanced chromatography and proprietary protocols for high purity, ensuring clean, safe, efficacious Mito-mRNA for sensitive applications.

3. Quality Control

   • Integrity Assessment: Gel electrophoresis and capillary electrophoresis are used to verify the full-length nature and absence of degradation.
   • Concentration Determination: Spectrophotometric analysis accurately quantifies mRNA yield.
   • Purity Analysis: Evaluation of dsRNA content, endotoxin levels, and other impurities to ensure safety and prevent off-target effects.
   • Sequence Verification: Sanger sequencing confirms the exact sequence, including the MTS, ensuring accurate protein translation.
   • Functional Validation (Optional): In vitro assays can be performed to confirm mitochondrial localization and expression of the target protein within relevant cell lines, providing empirical evidence of the mRNA's functional activity.

4. Final Deliverables

   • High-Purity Custom Mitochondrial Targeting mRNA: Optimized for your specific application.
   • Comprehensive Quality Control Report: Detailing mRNA integrity, concentration, purity, and other relevant parameters.
   • Sequence Verification Data: Confirmation of the accurate integration of your target gene and MTS.

5. Estimated Timeframe

   The typical timeframe for this service ranges from 2 to 5 weeks, depending on the complexity of the mRNA sequence, the specific modifications required, and the scope of quality control assays.

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What We Can Offer?

Customer Reviews

"Using Creative Biolabs' Custom Mitochondrial Targeting mRNA in our research has significantly improved our ability to study mitochondrial protein dynamics. The purity of their mRNA is exceptional, leading to highly specific targeting and robust expression, which was crucial for our sensitive in vivo experiments. We've seen a noticeable reduction in non-specific cellular responses compared to other providers."

2024, Dr. Chen

"Creative Biolabs' expertise in designing MTS for neuronal applications was a game-changer for our work on synaptic plasticity. Their Mito-mRNA allowed us to precisely deliver proteins to distal neuronal mitochondria, a challenge we struggled with for years. The detailed QC data provided immense confidence in our results, ensuring our findings were truly attributable to the targeted mitochondrial expression."

2023, Prof. Patel

"The Custom Mitochondrial Targeting mRNA Synthesis service from Creative Biolabs streamlined our pre-clinical drug screening for mitochondrial-related diseases. Their rapid turnaround and consistent quality enabled us to test multiple therapeutic candidates much faster than anticipated, providing a clear advantage in our competitive research landscape. Their team's understanding of therapeutic applications is truly valuable."

2024, Dr. Khan

FAQs

Sub-cellular fractionation: Fractionate cells into cytosolic, mitochondrial, and nuclear parts, then use RT-qPCR to detect mRNA in mitochondrial fractions, with higher levels indicating successful targeting.

Creative Biolabs' Custom Mitochondrial Targeting mRNA Synthesis service offers a precise and powerful tool for advancing mitochondrial research and developing targeted therapeutics. By enabling the direct expression of proteins within mitochondria, we empower scientists to explore complex biological questions and address critical unmet medical needs in areas ranging from neurodegeneration to cancer. Our commitment to quality, combined with our deep scientific expertise, ensures that you receive highly pure, functionally active mRNA tailored to your exact specifications.

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References

1. Criscuolo, Daniela, et al. "Targeting mitochondrial protein expression as a future approach for cancer therapy." Frontiers in Oncology 11 (2021): 797265. DOI: 10.3389/fonc.2021.797265. Distributed under Open Access license CC BY 4.0, without modification.