sgRNA Synthesis Service
Overview
CD BioGlyco specializes in providing high-quality, scalable single-guide RNA (sgRNA) synthesis crucial for all phases of CRISPR-Cas research and therapeutic development. As an essential component of the CRISPR-Cas system, the sgRNA dictates the specificity and efficacy of gene editing, making its quality paramount to experimental success. Leveraging over 20 years of experience, our service delivers custom sgRNA molecules engineered for maximum on-target activity and stability, minimizing off-target events and ensuring reproducible results across functional genomics, high-throughput screening, and cutting-edge preclinical investigations. We bridge the gap between complex molecular design and reliable, large-scale production, enabling our clients to accelerate their scientific timelines from discovery to clinical translation.
Core Technologies
Our expertise is rooted in deploying the optimal synthesis strategy tailored to your specific application, scale, and purity requirements. We offer a robust technology platform encompassing both rapid, cost-effective methods and advanced, chemically precise manufacturing processes.
- Advanced Chemoenzymatic Synthesis and Ligation
This high-end approach combines the precision of solid-phase chemical synthesis for the guide portion with enzymatic ligation to efficiently construct the full-length sgRNA scaffold. This method is particularly advantageous for therapeutic and in vivo applications, as it allows for the precise incorporation of multiple chemical modifications, resulting in highly stable, non-immunogenic, and potent guide molecules with minimal truncated species.
- Chemical Synthesis (Solid-Phase)
For applications demanding the highest standards of purity, chemical synthesis enables complete control over every phosphodiester bond. This process facilitates the systematic incorporation of stabilizing modifications, such as 2'-O-methyl and phosphorothioate linkages, which protect the RNA from nucleolytic degradation, a necessity for challenging delivery formats like lipid nanoparticles (LNPs).
- Optimized In Vitro Transcription (IVT)
We offer a highly optimized IVT service for projects requiring rapid turnaround or large-scale production for initial screening and functional genomics studies. Our proprietary template design and enzyme formulation enhance transcription efficiency and yield. We follow this with advanced purification protocols to significantly reduce the presence of abortive transcripts, ensuring high levels of full-length, active sgRNA suitable for RNP formation.
Precision-synthesized sgRNA for High-Efficiency Editing.
Our sgRNA synthesis service is designed to meet the diverse needs of the life science community, from academic research to commercial therapeutic development. Our scope covers comprehensive capabilities:
- Scale
Flexible production scales ranging from microgram quantities essential for small-scale functional assays, up to multi-gram quantities required for large-scale in vivo screens and clinical material production.
- Sequence Complexity
Synthesis of virtually any target-specific sequence for various Cas nucleases (e.g., Cas9, Cas12a, high-fidelity variants), including customized scaffold designs for enhanced editing specificity or novel fusion proteins.
- Chemical Modifications
Extensive catalog of stabilizing modifications, including 2'-O-methyl and phosphorothioate backbones to increase nuclease resistance and in vivo half-life, critical for successful systemic delivery.
- Formulation Options
Providing sgRNA in custom buffers, salt-free lyophilization for optimal delivery vehicle formulation, or pre-complexed as Ribonucleoprotein (RNP), ready for immediate transfection or microinjection.
- Comprehensive QC
Identity verified by mass spectrometry (MS), integrity assessed by gel or capillary electrophoresis, purity validated by HPLC, and trace contaminant detection (endotoxins, residual DNA/protein) suitable for the intended use.
Workflow
Design & Consultation
Expert bioinformatic analysis to identify and score optimal target sequences. We leverage proprietary algorithms to minimize off-target effects and maximize on-target activity, followed by a dedicated consultation to finalize sequence, modification strategy, and purity specifications.
Template Generation
Preparation of high-fidelity starting materials (linear DNA templates for IVT or oligonucleotide building blocks for chemical synthesis). This stage involves rigorous quality control of initial components to ensure successful downstream production.
Synthesis & Modification
Execution of the chosen synthesis method (IVT, chemical, or chemoenzymatic ligation). Chemical modifications are precisely introduced during this phase to enhance the sgRNA's stability, nuclease resistance, and cellular uptake properties.
Advanced Purification
Implementation of high-resolution purification techniques, including HPLC, ion-pair reversed-phase chromatography, or chromatography-based methods for therapeutic grade, to isolate the full-length sgRNA product and effectively remove truncated byproducts, unincorporated reagents, and transcription machinery components.
Rigorous Quality Control
Comprehensive analytical testing, including mass MS for identity confirmation, capillary electrophoresis (CE) or polyacrylamide gel electrophoresis (PAGE) for purity and integrity, and functional testing (optional) to confirm on-target Cas-binding and cleavage activity.
Delivery & Documentation
The final, high-purity product is quantified, packaged according to client specifications (e.g., lyophilized or in solution), and shipped alongside a detailed certificate of analysis (CoA) and full technical documentation for seamless integration into your research pipeline.
Publication Data
Journal: Nature Communications
DOI: 10.1038/s41467-025-61748-4
IF: 15.7
Published: 2025
Results: This study presents a novel split sgRNA strategy to enhance the functionality and application scope of the CRISPR-Cas12b system. The authors address a key limitation of Cas12b—its requirement for a long, single-guide RNA (sgRNA) exceeding 100 nucleotides—by engineering a modular system where the sgRNA is split into universal components (tracrDR or tracrRNA+DR) and a replaceable Spacer region. They demonstrate that this split configuration retains robust trans-cleavage activity comparable to full-length sgRNA while offering superior programmability. A significant innovation is the use of chemical modifications, such as glyoxal labeling or a photo-cleavable linker on the universal DR component, which allows for precise temporal control over Cas12b's nuclease activity. This enables compatibility with one-pot amplification assays like RPA, preventing unwanted degradation of the amplification template. The method was successfully applied to detect Epstein-Barr virus in clinical samples with sensitivity matching qPCR. Furthermore, the authors showcase a groundbreaking application by utilizing microRNAs themselves as the Spacer, enabling direct, amplification-free detection of specific microRNAs, which was validated in cell lines and patient plasma. This work significantly advances CRISPR diagnostics by providing a versatile platform for regulated nucleic acid detection and direct microRNA analysis.
Applications
Therapeutic Gene Editing and Drug Development
High-purity, chemically modified sgRNA is essential for developing novel therapies targeting genetic diseases, acting as a crucial component of in vivo and ex vivo gene correction strategies; our preclinical-grade material ensures low immunogenicity and optimal systemic delivery performance.
Functional Genomics and High-Throughput Screening (HTS)
Large, diverse libraries of sgRNA are used to systematically knock out, activate, or repress genes across the genome to identify novel drug targets, essential pathways, or synthetic lethal interactions in a massive parallel screening format.
Base and Prime Editing Systems
Our synthesis service supports specialized gRNA molecules required for next-generation CRISPR tools that perform precise single-base changes or small edits without introducing double-strand breaks, demanding exceptionally high fidelity and purity to prevent unwanted cutting.
Development of In Vivo Delivery Systems
Researchers formulating delivery vehicles, such as viral vectors (AAV, lentivirus), lipid nanoparticles (LNPs), or polymer conjugates, rely on our custom, high-concentration sgRNA products to optimize packaging, stability, and cell-specific targeting efficiency.
Advantages
Unparalleled Purity
We utilize proprietary multi-step chromatography, including robust HPLC/FPLC, ensuring the removal of truncated side products and contaminants which typically interfere with successful RNP formation and induce off-target effects. Our purity standards exceed 98%, essential for reliable in vivo studies.
Scalable Production
Our platform is engineered for seamless scalability, ensuring that the exact same high-quality sgRNA produced for initial validation can be manufactured at multi-gram quantities under strict quality management systems, significantly de-risking the transition to preclinical studies.
Enhanced Stability
Our expertise in synthetic and chemoenzymatic methods allows for the targeted incorporation of chemical modifications that dramatically boost the sgRNA’s stability and resistance to intracellular nucleases, resulting in longer cellular half-life and greater editing efficiency.
Minimization of Immunogenicity
By supplying highly purified, synthetic sgRNA, we inherently reduce the introduction of double-stranded RNA contaminants often present in standard IVT preparations, thereby mitigating unwanted innate immune responses in cell and animal models.
Frequently Asked Questions
Customer Review
We rely on CD BioGlyco for our high-throughput screening libraries. The yield and quality of the IVT sgRNA we receive are consistently superior to our internal production, minimizing failed screens and reducing the total cost of our functional genomics efforts. Their rapid, reliable service allows us to pivot quickly and test more targets than ever before.
— Manager D. Patel, Head of Genome Engineering, Industrial Biotechnology Unit
The chemically modified sgRNA from CD BioGlyco drastically improved the outcome of our in vivo mouse model experiments. The enhanced stability meant we could use lower, less toxic doses while maintaining high editing efficiency. The technical support team was instrumental in guiding our choice of modifications. The difference in data quality was immediately apparent.
— Professor A. Rodriguez, Department of Molecular Medicine
We needed a custom scaffold sgRNA for a novel Cas variant, and CD BioGlyco was the only provider capable of meeting the complex synthesis and purification demands. The material arrived on time, was perfectly synthesized, and integrated seamlessly into our AAV packaging protocol. Their expertise is truly high-end.
— Dr. J. Kim, Research Associate, Viral Vector Core
Associated Services
To help our clients maximize the potential of their custom sgRNA, we offer a suite of complementary services designed to streamline the entire RNA-based therapeutic and research pipeline.
RNA Production and Purification Process Development
Accelerate your time to clinic with our expertise in optimizing sgRNA (and other RNA species) manufacturing processes. This includes developing robust, high-yield, and scalable protocols and performing rigorous analytical method validation.
RNA-based Drug Formulation Development Service
We provide expert consultation and practical development services focusing on complex RNA delivery, including optimization of LNP formulation and encapsulation parameters for maximum sgRNA protection, stability, and in vivo targeting efficiency.
CD BioGlyco stands as your expert partner in cutting-edge gene editing. Our sgRNA synthesis service provides unparalleled quality, stability through advanced chemical modification, and scalable production, ensuring that your research—from fundamental discovery to clinical translation—is built on the most reliable foundation possible. Contact us to obtain the highest purity sgRNA available, empowering you to achieve exceptional editing efficiency and reproducible results.
Reference
- Wang, J.; et al. Regulating cleavage activity and enabling microRNA detection with split sgRNA in Cas12b. Nature Communications. 2025, 16(1): 6392. (Open Access)