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Prime editing (PE) provides enhanced precision compared to traditional Cas9 systems and offers a potential alternative for CRISPR-based gene and cell therapeutic development by eliminating the need for double-stranded DNA breaks, which are associated with off-target edits.
Prime editing guide RNA (pegRNA) is one of the two essential components for prime editing. It not only identifies the target site but also encodes the new DNA sequence for the edit. The pegRNA consists of a spacer, scaffold, reverse transcription template (RTT) containing the desired edit sequence, and a primer binding site (PBS) to initiate reverse transcription. The nickase-Cas9 (nCas9) fused with a reverse transcriptase is guided to the target site by the pegRNA, where it introduces a single-strand break (or "nick") in the target DNA. The pegRNA then initiates reverse transcription to replace the removed sequence with the new template sequence.
The complex design of pegRNA often results in lengths exceeding 110 nucleotides, making synthesis challenging. Leveraging over 22 years of RNA oligo synthesis expertise, we proudly offer desalted and HPLC-purified pegRNA up to 266 nucleotides in length with default 2'-O-methyl + phosphorothioate modifications for enhanced stability and performance.
| Length* | Quantity | Desalt Grade | HPLC Grade |
|---|---|---|---|
| 110-150 nt | 2 nmol | $240 | Inquire for pricing |
| 5 nmol | $330 | ||
| 10 nmol | $450 | ||
| 50 nmol | $1100 | ||
| 100 nmol | $1600 | ||
| 151-266 nt | 2-100 nmol | Inquire for pricing |
GenScript offers INDEdit and cGMP-grade products for IND submissions and clinical research. Learn more.
Need help from an expert?
pegRNA design tool - Empower precise and easy design
Already have your pegRNA design? Use our simple order form to order or get a quote:
Shawn Yang
Prime Medicine
I have been using GenScript to order customized critical reagents for an important assay. GenScript can always deliver materials efficiently with good quality. Their technical support is great at addressing any additional questions.
Up to 266 nt for Both Desalt &
HPLC Purified Grades
Precise molecular weight, high purity, excellent batch-to-batch consistency
Proven Editing Efficiency
Editing efficiency over 60%
Modifications to enhance efficiency
One-Stop Solution
PE2/PE3 mRNA available
Validated experiment protocol
RUO to cGMP Capability
Supporting research to IND application & clinical trials
| Service | pegRNA (Prime Editing guide RNA) Synthesis Service |
|---|---|
| Length | 110-266 nt* *For <110 nt or>200 nt RNA oligo synthesis, please inquire here |
| Modification |
|
| Purification | Desalt & HPLC purification options (Purity is guaranteed ≥ 85% for HPLC purified grade )* |
| Quantity | 2-100 nmol, lyophilized powder. Optional suspension in TE buffer or NF-dH2O also available. For other quantities, please inquire here |
| QC Reports | COA and MS report (HPLC report available for HPLC purity grade) |
| Production Time | 7 business days (Excluding shipping time) |
* ≥85% purity is guaranteed for HPLC-purified pegRNA 110-200 nt in length. For other lengths, please inquire here.
Need help from an expert?
Prime Editing Products: Rapid and Efficient Validation and Optimization of your PE Editing System
Are you looking to validate your prime editing experimental system or optimize for editing efficiency? Try our off-the-shelf Prime Editing products:
(Refer to the protocol here and see validation data in Case 2 here.)
Your search returns no product results. Please explore our custom peptide synthesis solutions.
Please download the datasheet for detailed information about the sequence and other specifications.
HPLC purified pegRNA (266 nt): Precise molecular weight
Desalt purified pegRNA (218 nt): Precise molecular weight
HPLC purified pegRNA (266 nt): Purity up to 94.86%
Methods: HEK293T cells (0.2 million cells) were transfected with 1 μg PE2/PE3 mRNA, 30pmol nicking sgRNA and 90pmol pegRNA of different lengths (HPLC grade) targeting the HEK3 gene, via electroporation. Genome DNA was then extracted 3 days post-transfection for editing efficiency analysis via Sanger sequencing.
Results: pegRNA with a tevopreQ1 motif achieved editing efficiencies as high as 43.4%, which was significantly higher than that of the pegRNA without the tevopreQ1 motif.
Note: the 218 nt pegRNA is an extended version of 181 nt with a tevopreQ1 motif (37 nt) at the 3' end. It has been reported that adding the RNA structural motifs evopreQ1 or mpknot to the 3' end of a pegRNA can protect the 3' end from exonuclease digestion (Nelson, J. W. et al., 2022).
Methods: 0.2 million HEK293T cells were transfected with different dosages of 141 nt long pegRNA (desalt or HPLC purified grade) targeting the HEK3 gene, via electroporation. Nicking sgRNA was also added in a ratio of 3:1 to pegRNA. PE2/PE3 mRNA was added with increasing dosages from 0.15 μg, 0.25 μg, 0.375 μg, 0.5 μg, 0.75 μg to 1.0 μg. Genome DNA was then extracted 3 days post-transfection for editing efficiency analysis via Sanger sequencing.
Results: Editing efficiencies up to 52.5% for desalt and 63.5% for HPLC-purified pegRNA was achieved. Editing efficiency increased in a dose-dependent manner.
Methods: HEK293T cells (0.2 million cells) were transfected with a 1 μg pCMV-PE2/PE3 plasmid and 45pmol of 141 nt long pegRNA targeting the HEK3 gene, via electroporation. Genome DNA was then extracted 3 days post-transfection for editing efficiency analysis via Sanger sequencing. 15pmol of additional nicking sgRNA was added to direct the prime editor enzyme to nick the non-edited strand in the PE3 system.
Results: GenScript's HPLC grade and desalt pegRNA achieved comparable editing efficiency to Vendor I at a more cost-effective price.
97 to 103 nt, improving your editing efficiency at a cost-effective price
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40 to 49 nt, with superior quality and editing efficiency up to 98%
Learn MoreGenScript can also synthesize desalt and HPLC purified guide RNA compatible with saCas9, Cas12f/CasMINI, Cas13, and other Cas nuclease variants.
Click here to enter your guide sequence (target sequence + scaffold sequence) and order online.
Prime Editing Protocol for HEK293T cells with Thermo Fisher Neon® Transfection System
Free Download
A Beginner's Guide to Prime Editing Guide RNA (pegRNA)
Free Download
