Prime Gene Editing Service

Prime Editing (PE) is a groundbreaking 'search-and-replace' gene editing technology that significantly expands the scope of programmable genomic modification. Unlike traditional CRISPR/Cas9 which relies on error-prone double-strand break (DSB) repair, or Base Editing which is limited to specific base conversions, Prime Editing utilizes a fusion protein—a Cas9 nickase (nCas9) fused to a reverse transcriptase (RT)—guided by a specialized Prime Editing guide RNA (pegRNA).

This innovative system allows for the precise introduction of virtually all 12 types of point mutations, as well as targeted small insertions and deletions (indels), all without creating a DSB or requiring a separate DNA donor template. CD Biosynsis provides comprehensive Prime Editing CRO services, leveraging optimized PE platforms and proprietary pegRNA design algorithms to deliver high-fidelity, complex genetic modifications in various cell types, accelerating your discovery and therapeutic development programs.

Highlights

Our Prime Editing platform offers unparalleled flexibility and precision for challenging genetic modifications:

• Universal Edit Types: Achieve all 12 possible point mutations , and precise small insertions (up to ~44 bp) or deletions (up to ~80 bp) , surpassing the limitations of Base Editing and standard CRISPR.
• DSB-Free Precision: By utilizing a Cas9 nickase, PE avoids the formation of DNA Double-Strand Breaks, drastically minimizing the risk of unwanted large deletions, rearrangements, and non-homologous end joining (NHEJ) byproducts.
• Optimized PE Components: We employ the latest generation of Prime Editors (PE2, PE3, etc.) and proprietary pegRNA/ngRNA designs to maximize editing efficiency and target specificity across diverse genomes.
• Advanced Bioinformatics: Rigorous computational and wet-lab screening protocols for pegRNA design ensure optimal target selection and minimize off-target editing, critical for therapeutic applications.

Applications

Prime Editing's versatility makes it the ideal tool for complex genomic engineering in both research and clinical contexts:

Precision Disease Modeling

           

Accurate creation of patient-relevant single-nucleotide polymorphisms (SNPs) and complex small indels in iPSCs or primary cells for disease mechanism studies.

Therapeutic Gene Correction

Direct correction of pathogenic mutations (point mutations, small duplications/deletions) in therapeutically relevant cells for cell and gene therapy development.

Functional Gene Tagging/Reporter Insertion

Precise insertion of short sequences for epitope tags, fluorescent reporters (e.g., GFP) or drug-selection markers at specific genomic loci.

High-Throughput Functional Screening

Generation of diverse, high-complexity libraries containing precise point mutations to map functional domains and regulatory element impact.

Solutions

CD Biosynsis offers a full spectrum of Prime Editing services, from initial design to validated clonal cell delivery.

Advanced pegRNA/ngRNA Design & Synthesis

Bioinformatic-driven design of optimal Prime Editing guide RNA (pegRNA) and nicking guide RNA (ngRNA) sequences for maximum efficiency.

Engineered Prime Editor Expression Constructs

Construction and optimization of PE plasmids, mRNA, or RNP components utilizing the most effective PE variants (PE2/PE3/PE4) for your specific cell line.

Complex Cell Line & iPSC Engineering

Generation of precisely edited stable cell lines, including difficult-to-edit primary cells and induced pluripotent stem cells (iPSCs).

Optimized Delivery and Screening

Selection and optimization of delivery methods (lipofection, electroporation, viral vectors) followed by high-throughput clonal screening.

Deep Sequencing Validation & QC

Comprehensive next-generation sequencing (NGS) to confirm on-target edit fidelity, homozygosity, and assess potential off-target effects.

Workflow

Our structured Prime Editing workflow ensures maximum success rates and high-quality deliverables:

• Project Consultation & Design: Define the exact edit (point mutation, insertion, or deletion). Our experts design and screen multiple pegRNA and ngRNA sequences, selecting the optimal Prime Editor version (e.g., PE2, PE3) based on the target locus.
• Component Generation & Optimization: Synthesis and validation of high-quality PE plasmids/mRNA and pegRNA/ngRNA. We optimize the ratios and delivery conditions for your specific cell line to maximize the window for successful editing.
• Cell Line Engineering & Clonal Isolation: Deliver the optimized PE components into the host cells. Utilize FACS or drug selection (if applicable) followed by single-cell plating and expansion of presumptive positive clones.
• Rigorous Quality Control & Validation: Perform comprehensive sequencing (Sanger and/or NGS) on isolated clones to confirm the presence and fidelity of the precise Prime Edit. Crucially, we screen for any unintended byproducts (such as DSB-mediated indels).
• Final Delivery & Documentation: Deliver the validated, cryopreserved edited cell line/model. A complete data package includes a detailed QC report, full sequencing data, and the optimized protocol for future use.

We are committed to delivering reproducible and clinically-translatable results. Every project includes:

• Guaranteed Precision: Confirmation of the exact desired edit without unintended byproducts (low DSB-mediated indel rate).
• Full Data Transparency: Comprehensive raw and analyzed NGS data to prove precise point mutation, insertion, or deletion.
• Technical Expertise: Access to our dedicated project managers who are Prime Editing experts, providing timely technical support and project updates.
• Proprietary Optimization: Utilization of internally validated PE system variants and delivery protocols tailored for difficult targets.