HPLC purified pegRNA (218 nt): Precise molecular weight
Desalt purified pegRNA (218 nt): Precise molecular weight
HPLC purified pegRNA (218 nt): Purity up to 92.78%
Prime Editing (PE) offers unprecedented precision compared to traditional Cas9 systems and serves as a safer alternative for CRISPR-based gene and cell therapeutic development by eliminating the need to create double-stranded DNA breaks, which can result in 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 is composed of a spacer, scaffold, reverse transcription template (RTT) with the desired edit sequence, and a primer binding site (PBS) for initiating reverse transcription. The nickase-Cas9 (nCas9) fused with a reverse transcriptase is guided to the target site by the pegRNA, where it makes a single-strand break (or ‘nick’) in the target DNA. Subsequently, the pegRNA initiates reverse transcription to replace the removed sequence with the new template sequence.
The intricate design of pegRNA often results in lengths exceeding 110 nt, making synthesis challenging. Leveraging our 22+ years of expertise in RNA oligo synthesis, we are proud to offer desalt and HPLC-purified pegRNAs up to 231 nt in length, along with over 200 modification options to meet your research needs.
*Promotions exclude customers in China, Japan, France, Spain, Norway, Switzerland, Italy, Sweden, Denmark, Finland, Portugal, Iceland, Monaco and some organizations. Valid for orders placed between Sep 3rd and Dec 31st 2024 only and cannot be applied to existing orders.
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 to address any additional questions.
Up to 231 nt for Both Desalt &
HPLC Purified Grades
Precise molecular weight, high purity, excellent batch-to-batch consistency
Proven Editing Efficiency
Editing efficiency over 60%
Modification to enhance efficiency
One-Stop Solution
PE2/PE3 mRNA available
Validated experiment protocol
RUO to cGMP Capacity
Supports research to IND application & clinical trials
| Service | pegRNA (Prime Editing guide RNA) Synthesis Service |
| Length | 110-231 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) |
* Purity is guaranteed ≥ 85% for HPLC purified grade for pegRNAs within 200 nt, other length please inquire here
Need help from an expert to support your pegRNA request?
| Length* | Quantity | Desalt Grade | HPLC Grade |
|---|---|---|---|
| 110-150 nt | 2 nmol | $240 $168 | Inquire for pricing |
| 5 nmol | $330 $231 | ||
| 10 nmol | $450 $315 | ||
| 50 nmol | $1100 | ||
| 100 nmol | $1600 | ||
| 151-231 nt | 2-100 nmol | Inquire for pricing |
HPLC purified pegRNA (218 nt): Precise molecular weight
Desalt purified pegRNA (218 nt): Precise molecular weight
HPLC purified pegRNA (218 nt): Purity up to 92.78%
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 with more cost-effective price.
