Comprehensive Overview of Prime Editing Guide RNA (pegRNA) Synthesis
Precisio n genome editing has revolutionized biological research and therapeutic development. At BOC Sciences, we provide high-quality Prime Editing Guide RNA (pegRNA) Synthesis Services to facilitate precise genome modifications. Our advanced synthesis capabilities ensure high-purity, sequence-verified pegRNAs, optimizing their stability and functionality for various research and industrial applications. With extensive expertise in RNA synthesis and genome engineering, we offer custom pegRNA synthesis services tailored to researchers' specific experimental requirements. Whether your goal is gene correction, targeted mutagenesis, or precise base conversion, our pegRNA synthesis solutions ensure reliability, efficiency, and scalability.
How do you submit a request for prime editing guide rna synthesis?
To initiate your pegRNA synthesis request, please follow these steps:
- Submit the sequence details: Provide the target sequence, desired edit, and PBS/RT template sequence (design assistance is available).
- Specify modifications: Indicate any desired chemical modifications for stability or delivery.
- Define your application: State whether it's for in vitro, in vivo, or gene therapy research.
- Indicate quantity and purity: Choose research-grade or large-scale synthesis with your preferred purification method.
- Place your order: Submit your request via the BOC Sciences online portal for a seamless process.
What is Prime Editing?
Prime editing is an advanced genome-editing technology that allows precise modifications without inducing double-strand breaks (DSBs). Unlike traditional CRISPR- Ca s 9, which relies on error-prone repair pathways, prime editing uses a fusion of a catalytically impaired Ca s 9 (nCa s 9) and a reverse transcriptase (RT) to introduce targeted genetic changes with minimal off-target effects. Compared to traditional genome-editing technologies, prime editing offers several advantages:
- Greater precisio n: Enables targeted insertions, deletions, and point mutations without generating DSBs.
- Broad applicability: Compatible with diverse cell types and organisms.
- Reduced off-target risks: Minimizes unintended genomic modifications.
- Expanded editing scope: Overcomes limitations of existing base-editing and CRISPR- Ca s 9 techniques.
By selecting the appropriate vector backbone, promoter, and delivery mechanism, sgRNA vectors can be optimized for different cell types, gene targets, and experimental conditions.
What is Prime Editing Guide RNA?
Prime Editing Guide RNA (pegRNA) is a specialized RNA molecule that directs the prime editing process. Unlike traditional sgRNA used in CRISPR- Ca s 9, pegRNA has an extended structure, incorporating:
- Spacer sequence – Guides the Ca s 9 variant to the target genomic site.
- Primer-binding site (PBS) – Serves as an anchoring sequence for the reverse transcriptase.
- Reverse transcriptase template (RTT) – Encodes the desired genetic modification.
The unique structure of pegRNA allows the precise rewriting of genetic sequences without requiring a donor DNA template, making it a critical component in prime editing experiments.
High-Purity pegRNA Synthesis Services for Precisio n Genome Editing
BOC Sciences provides a comprehensive and customizable Prime Editing Guide RNA (pegRNA) Synthesis Service, designed to support researchers in achieving highly precise genome modifications. Our expertise in RNA chemistry, synthesis platforms, and sequence optimization allows us to deliver pegRNAs with enhanced editing efficiency, stability, and specificity.
Selection of pegRNA Synthesis Methods
BOC Sciences utilizes multiple state-of-the-art RNA synthesis approaches, allowing for flexibility in scale, purity, and structural complexity:
(1) Solid-Phase Phosphoramidite Synthesis
- Best for short-to-medium-length pegRNAs with high purity.
- Compatible with various chemical modifications for enhanced stability.
(2) Enzymatic Transcription (IVT, In Vitro Transcription)
- Ideal for longer pegRNAs (>100 nt) that are challenging to synthesize chemically.
- Enables scalable production for high-throughput applications.
- Optionally includes cap structure and poly(A) tails for in vivo applications.
(3) Hybrid Synthesis (Chemical + Enzymatic)
- Used for generating high-fidelity pegRNAs with both precise chemical modifications and long RNA sequences.
- Provides flexibility in pegRNA customization while maintaining high synthesis efficiency.
Selection of pegRNA Types
We offer pegRNAs tailored for different prime editing applications, ensuring maximum efficiency and reliability.
(1) Standard pegRNA
- Includes a guide RNA (gRNA), a reverse transcriptase (RT) template, and a primer-binding site (PBS).
- Designed for standard prime editing experiments requiring high specificity.
(2) Dual pegRNA Systems
- Used for multiplexed editing or large insertions/deletions.
- Allows for simultaneous modifications at multiple loci.
(3) pegRNA Variants with Secondary Structure Optimization
- Custom-engineered to enhance stability and efficiency in challenging genomic contexts.
- Designed to reduce RNA degradation and off-target effects.
Selection of Chemical Modifications of pegRNA
Chemical modifications can significantly enhance the stability, durability, and performance of pegRNAs, particularly in cell-based and in vivo applications. BOC Sciences provides:
(1) 5' and 3' End Modifications
- 5′ Cap structures for improved stability.
- 3′ Poly(A) tails to enhance RNA longevity.
- Phosphorothioate (PS) linkages to increase resistance to nucleases.
- 2′-O-Methyl (2′-OMe) and 2′-Fluoro (2′-F) substitutions to reduce immune activation and increase resistance to RNases.
- Locked Nucleic Acids (LNAs) to enhance hybridization efficiency and increase pegRNA half-life.
(3) Modified Bases for Enhanced Editing Efficiency
- Pseudouridine (Ψ) and N1-methylpseudouridine (m1Ψ) for improved RNA stability and reduced immunogenicity.
- 5-Methylcytidine (5mC) to modulate RNA folding and stability.
Custom Length & Yield Options of pegRNA
Our synthesis service accommodates a wide range of pegRNA lengths and production scales, ensuring flexibility for different research applications.
(1) pegRNA Length Options
- Customizable lengths ranging from 80 nt to over 300 nt, depending on sequence complexity and editing requirements.
- Long pegRNAs are synthesized using high-yield enzymatic transcription with rigorous quality control.
(2) Production Scale
- Small-scale synthesis for pilot studies and proof-of-concept experiments.
- Medium-to-large-scale synthesis for high-throughput screening and industrial applications.
- Bulk synthesis for therapeutic and preclinical research requiring high RNA quantities.
Step-by-Step Workflow of pegRNA Synthesis and Optimization
Our synthesis process follows a stringent workflow to ensure high-quality, fully functional pegRNAs:
01
Sequence Design Consultation
- Bioinformatics-driven optimization of pegRNA sequences.
- Custom PBS and RT template design tailored to specific targets.
02
Custom pegRNA Synthesis & Chemical Modifications
- High-purity pegRNA synthesis using advanced solid-phase synthesis techniques.
- Optional modifications for improved RNA stability and intracellular performance.
03
Purification & Quality Control
- HPLC purification and PAGE analysis for high-purity pegRNAs.
- Mass spectrometry and UV spectrophotometry for sequence verification.
04
Functional Validation & Delivery
- Optional in vitro functional assays to confirm editing efficiency.
- Lyophilized or solution-based pegRNA formats for easy experimental integration.
With this systematic and quality-driven workflow, BOC Sciences guarantees highly efficient and precisely designed pegRNAs that empower researchers to push the boundaries of genome editing.
Key Advantages of Choosing Our pegRNA Synthesis Service
BOC Sciences is a trusted leader in RNA synthesis and genome editing solutions, offering unparalleled expertise and innovation in pegRNA production.
Expertise in RNA Engineering
With decades of experience in RNA synthesis, we provide optimized pegRNAs tailored for superior prime editing efficiency.
High-Quality, Purified Products
Rigorous quality control ensures high-purity and high-yield pegRNA synthesis, minimizing unwanted byproducts and degradation.
Custom Modifications for Stability & Delivery
Options for chemical modifications to enhance RNA longevity, reduce immunogenicity, and improve intracellular stability.
Scalable & Cost-Effective Solutions
From small-scale research projects to large-scale industrial production, we accommodate diverse project needs with competitive pricing.
Fast Turnaround & Reliable Support
Rapid synthesis timelines with dedicated technical support for project-specific requirements.
Key Applications: Expanding Research Possibilities with pegRNA
Precisio n Gene Therapy Development
- Correction of pathogenic mutations in genetic diseases such as cystic fibrosis, sickle cell anemia, and Duchenne muscular dystrophy.
- Development of ex vivo and in vivo gene therapies that introduce precise genetic corrections in patient-derived cells or directly in tissues.
- Enhanced safety and efficacy compared to traditional gene-editing approaches, due to the absence of double-strand breaks (DSBs).
Functional Genomics & Disease Modeling
- Introduction of precise knock-ins, deletions, and point mutations to study gene function and regulatory mechanisms.
- Generation of disease-specific mutations in cell lines to model human genetic disorders.
- Development of reporter constructs and tagged proteins to study protein interactions and localization.
Cell Line & Animal Model Engineering
- Creation of genetically modified cell lines with targeted modifications for research and drug screening applications.
- Generation of transgenic and knock-in/knock-out animal models with precise genetic edits for biomedical research.
- Establishment of patient-derived disease models to facilitate personalized medicine research.
Drug Discovery & Synthetic Biology
- Development of genetically engineered cell lines optimized for drug screening, target validation, and pharmaceutical production.
- Synthetic biology applications, including the engineering of bacteria and yeast for biofuel production, biomaterials, and industrial bioprocesses.
- Tailored modifications in cellular pathways to optimize protein expression, metabolic engineering, and therapeutic molecule production.
FAQs about Our pegRNA Synthesis Service
Can pegRNA be delivered in different formats?
Yes, we provide pegRNA in lyophilized or solution format, tailored to your experimental needs. Custom buffer options are also available.
Do you provide pegRNA controls for experimental validation?
Yes, we offer positive and negative control pegRNAs to help validate prime editing efficiency and optimize experimental conditions.
Can pegRNA be stored for long-term use?
Yes, pegRNA should be stored at -80°C for long-term stability. We also provide recommendations on storage buffers to maintain RNA integrity.
What is the typical length of pegRNA?
pegRNA typically ranges from 120 to 200 nucleotides, depending on the design. Our team optimizes the length based on editing requirements.
What is the recommended transfection method for pegRNA?
pegRNA can be delivered via electroporation, lipid-based transfection, or viral vectors, depending on the cell type. We provide guidance on delivery strategies.
Can you provide large-scale pegRNA synthesis for screening applications?
Yes, we support high-throughput pegRNA synthesis for genome-wide screens, functional genomics studies, and large-scale prime editing experiments.
At BOC Sciences, we are committed to delivering high-quality, customizable, and efficient pegRNA synthesis services to support cutting-edge genome-editing research. Contact us today to discuss your pegRNA synthesis needs and accelerate your prime editing projects with confidence!
