GMP sgRNA-IVT
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.
GMP sgRNA IVT Manufacturing Platform
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.
GMP sgRNA IVT Highlights
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GMP sgRNA IVT Advantages
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Cost-effective
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Safer, no organic chemicals
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High gene editing efficiency
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Client-Centric Partnership
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Flexible and tailored the processes to accommodate your needs
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Proactive scientific and regulatory support
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Optimized GMP sgRNA Manufacturing Process
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Streamlined upstream and downstream processes
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Scalable manufacturing process, up to grams of product
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High purity sgRNA, >90%
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Excellence in Quality
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Robust QMS system
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Full spectrum in-house quality testing
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Compliance with FDA/EMA/NMPA requirements
GMP sgRNA IVT Manufacturing Workflow
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.
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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.
CRISPR RNP Formulation Service
sgRNA 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.
High Quality GMP sgRNA Produced
Fig. 1. Purity analysis of sgRNA by HPLC
Fig. 2. The purity of sgRNA by agarose gel electrophoresis
High Quality GMP sgRNA Produced
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.
GMP sgRNA QC Assays
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.
Test | Test Methods |
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Appearance | Visual method |
RNA Concentration | UV absorbance |
Purity | HPLC, A260/280 |
Identifying Sequence | Sanger sequencing |
Molecular Weight | Mass spectrometry |
Endotoxin Detection | BDBU - GEL-CLOT Method |
Sterility | Culture method |
Protein Residuals | Qubit / MicroBCA |
pH | pH meter |
DNA Residuals | qPCR |
dsRNA | ELISA |
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Why is a three-tiered cell bank required?According to regulations, a complete culture cell bank must be established before GMP-grade plasmid production, including the primary cell bank, the master cell bank and the working cell bank.
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What is the storage temperature for plasmids? How long can they be stored?According to BriGene’s many years of GMP plasmid production experience, the storage temperature of plasmids is -20°C, and the long-term storage stability data can be traced back for more than half a year.
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Which strain is used to produce plasmids?uBriGene uses DH5α or Stbl3 for plasmid production, depending on the customer's choice.
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Why are plasmids modified for kanamycin resistance?Regulations require that the vectors used in gene therapy drugs must not introduce resistance to penicillin.