uBriGene's single guide RNA (sgRNA) in vitro (IVT) transcription, sgRNA IVT, offers the fastest, most cost-effective path to clinical gene editing proof-of-concept results.
Our GMP sgRNA IVT platform uses in vitro transcription to produce sgRNA without organic solvents, reducing large-scale production costs. Additionally, a patented aptamer sequence at the 3' terminus enhances sgRNA stability and boosts gene editing efficiency.
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The DNA template used for sgRNA IVT is double-stranded DNA generated from PCR. Our plasmid manufacturing platform can provide the GMP-grade plasmids used for sgRNA IVT manufacturing. Simply provide us with your target specific guide RNA sequence, and we will deliver GMP-grade sgRNA, ready for your CGT programs.
The in vitro transcribed crude sgRNA will be further purified via chromatography and sterile filtration, followed by aseptic fill and finish. We have established a GMP-compliant production process platform and a comprehensive quality control platform to meet the regulatory requirements of FDA, EMA, and NMPA.
The advantages of membrane matrix in downstream chromatography for plasmid purification include requiring only 30% of the time compared to conventional resin. Therefore, plasmid DNA is manufactured faster and at a much lower cost.
The lysates are clarified through scalable, fully closed depth filtration, followed by a cost-effective two-step purification process using anion exchange (AEX) and hydrophobic interaction chromatography (HIC). For larger-scale production membrane matrix chromatography allows for higher flow rates.
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Request a quotesgRNA complexes with Cas enzymes to form the ribonucleoprotein (RNP) complex which catalyzes CRISPR-mediated gene editing. uBriGene also has off-the-shelf GMP-grade Cas proteins and mRNA encoding Cas9, Cas12a, Cas13a, and Cas14a. We can provide RNP formulation services with your gene specific sgRNA. Request RNP Formulation.
The GMP-grade sgRNA IVT will be tested extensively for identity, purity, and safety using qualified and compendial methods to ensure its quality and safety for clinical use.
| Tests | Test Methods |
|---|---|
| Appearance | Visual method |
| RNA Concentration | UV absorbance |
| Purity | HPLC, A260/280 |
| Molecular Weight | Mass spectrometry |
| Identifying Sequence | Sanger sequencing |
| Endotoxin Detection | BDBU GEL-CLOT Method |
| Sterility | Culture method |
| Protein Residuals | Qubit / MicroBCA |
| pH | pH meter |
| DNA Residuals | qPCR |
| dsRNA | ELISA |
*Outsourced testing
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Fig. 1. Purity Analysis of sgRNA by HPLC.
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Fig. 2. The Purity of sgRNA by agarose gel electrophoresis.
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Fig. 3. Gene editing efficiency comparison of sgRNA IVT and chemically synthesized in iPSC cells. sgRNA and Cas9 RNPs targeting GOI 1 and GOI2 were transfected into cells. Gene knockout efficiency was determined by sequencing.
Guide RNA is a critical component in CRISPR-mediated gene editing, which is broadly utilized in cell and gene therapy clinical programs. sgRNA , is comprised of crRNA and tracrRNA. Currently sgRNA is produced via chemical synthesis. Our sgRNA IVT is more scalable and cost-effective.
The sgRNA binds with Cas enzymes to form a ribonucleoprotein (RNP) complex that drives CRISPR-mediated gene editing. The key benefits of CRISPR RNP include minimized off-target effects and the absence of DNA integration, resulting in enhanced gene editing safety.
uBriGene also has off-the-shelf GMP-grade Cas proteins and mRNA encoding Cas9, Cas12a, Cas13a, and Cas14a. We can provide RNP formulation services with your gene specific sgRNA. Request RNP Formulation.
Our in vitro transcribed guide RNA will be resuspended in nuclease-free water at 100 µM concentration or tailored to your specific requirements.
We offer both research grade and GMP sgRNA IVT, supporting CRISPR gene editing from proof of concept to clinical applications.
No. You do not need to include the PAM site in your gRNA design, as it is located next to the guide RNA targeting sequence in the genome.
5+ Batches of GMP sgRNA IVT have been successfully manufactured. The GMP sgRNA process is cost-effective and scalable for large scale clinical applications.
The aptamer at the 3’ end of the sgRNA ensures longer stability of the gRNA inside the cells and enhances CRISPR gene editing efficiency.
TuBriGene has solved the cytotoxicity and stability challenges of sgRNA in vitro transcription. Our IVT sgRNA production has proved to give higher gene editing efficiency than chemically synthesized guide RNA.
The estimated turnaround time for GMP-grade sgRNA IVT manufacturing is around 2 weeks.
Yes, sgRNA IVT performs well for both Indel gene knockout without donor DNA and homologous recombination with donor sequences.
Yes, we have Cas proteins and mRNAs, available both in research grade and GMP grade. We have Cas9, Cas12a, Cas13a, and Cas14a protein and mRNA products. Please view details of gene editing products. We can provide CRISPR RNP and LNP complexes with your specific sgRNA.
Yes, we do offer plasmid production services. We can provide circular plasmids, linear DNA, and closed-end linear DNA (for increased stability inside the cells). Please view details at GMP plasmid manufacturing.
End-to-end CDMO services with streamlined timelines and cost-effective processes.
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Ask the plasmid DNA expert! With over 300 GMP batches in our track record and our cost-effective platform technology, we can help accelerate your clinical programs.
