Cas9 mRNA
Wild type and SpyFi™ high-fidelity mRNA
Maximize editing efficiency with our optimized Cas9 mRNA—available in wild type and high-fidelity variants to reduce off-target effects. We accelerate your timelines with scalable, cost-effective solutions and expert technical support while providing continuity from discovery to translational and therapeutic development -GMP version coming soon! - accelerating program timelines and simplifying your CMC process.
Ordering
- Strategic collaboration between IDT and Aldevron delivers high-quality, scalable Cas9 mRNA solutions for gene editing applications
- Cas9 mRNA with high editing efficiency across diverse cell types using electroporation or lipid nanoparticles (LNP), benchmarked against a leading competitor
- High-fidelity Cas9 variant minimizes off-target gene editing for improved precision
- Simplified licensing with enzymatic capping
- Research Use Only (RUO) and CGMP-grade Cas9 mRNA (coming soon) with consistent performance—no process development required
- Low batch-to-batch and lot-to-lot variability ensures reproducibility and reliability
- Cas9 mRNA co-developed by IDT and Aldevron, combining CRISPR enzyme engineering and mRNA manufacturing expertise
Don’t see what you’re looking for?
Have questions for our CRISPR experts? Interested in bulk protein or a custom solution? Your time is valuable and we’ll prioritize your inquiry. Click on “Request a consultation” to provide brief information about your project, and we’ll be in touch to discuss it ASAP.
Request a consultationProduct Details
Why choose Cas9 mRNA from IDT and Aldevron?
- Flexible Cas9 mRNA formats: Wild type and SpyFi high-fidelity variants support a wide range of gene editing workflows
- Rapid delivery: Ships next day to your lab for faster experimental turnaround
- Validated editing performance: SpyFi Cas9 shows high on-target efficiency and low off-target effects across multiple cell lines and delivery methods, confirmed by IDT’s rhAmpSeq analysis system
- Optimized for human expression: Features NLS, 5’/3’ UTRs, codon optimization, and uridine depletion for enhanced translation
- Simplified licensing: 5’ enzymatic capping reduces licensing burden
- Stable and reliable: >100 nt poly(A) tail and high 5’ capping efficiency improves mRNA stability and expression consistency
- High purity: with low exogenous RNA present
- Trusted collaboration: Developed by IDT and Aldevron, combining CRISPR enzyme engineering and mRNA manufacturing excellence
This isn’t just another reagent—it’s a precision-engineered Cas9 mRNA solution born from the strategic partnership between IDT and Aldevron. Designed to accelerate discovery and simplify therapeutic development, it sets a new benchmark for fidelity, performance, and scalability in gene editing.
Figure 1: Introduce ready-to-use Cas9 mRNA together with sgRNA into your cells via electroporation, or liquid nanoparticle (LNP delivery) for CRISPR/Cas9 gene editing
Product Data
On- and off-target editing in primary T- cells
Figure 2. On-target editing at clinically relevant research sites and on- and off-targeting at HBB. Primary T-cells were thawed and activated 72 hours prior to nucleofection. Cas9 mRNA (1 µg) and sgRNA (2 µM) targeting AAVS1, B2M, CIITA, CISH, PD1 and TGFBR2 were delivered by nucleofection into T-cells using the Lonza Nucleofection™ System (A). Genomic DNA (gDNA) was extracted 72 hours post nucleofection and editing was assessed by NGS. Cas9 mRNA (1 µg) and sgRNA (2 µM) targeting HBB was delivered by nucleofection into T-cells using the Lonza Nucleofection™ System (B). The gDNA was extracted 72 hours post nucleofection and editing was assessed by NGS.
On- and off-target editing in immortalized HEK293 cells
Figure 3. On- and off-target editing in immortalized HEK293 cells. Cas9 mRNA (1 µg) and sgRNA (2 µM) targeting 12 sites in the HPRT locus were delivered by nucleofection into HEK293 cells using the Lonza Nucleofection™ System (A). Genomic DNA (gDNA) was extracted 72 hours post nucleofection and editing was assessed by NGS. Cas9 mRNA (1 µg) and sgRNA (2 µM ) targeting EMX1 was delivered by nucleofection into HEK293 cells using the Lonza Nucleofection™ System (B). The gDNA was extracted 72 hours post nucleofection and editing was assessed by NGS. Cas9 mRNA (0.125 µg) and sgRNA (2 µM) targeting EMX1 was delivered by nucleofection into HEK293 cells using the Lonza Nucleofection™ System (C). The gDNA was extracted 72 hours post nucleofection and editing was assessed by NGS.
Modified Cas9 mRNA shows reduced immune response vs unmodified and competitor mRNAs
Figure 4. IFN Response generated by THP-1 dual cells transfected with modified and unmodified Cas9 mRNAs. THP-1 dual cells were transfected in sextuplicate with 100 ng of the indicated mRNAs complexed with 1 µL transfection reagent Lipofectamine™ MessengerMAX™. After 24 hours, Lucia expression in the media was assayed as a measure of IFN activity.
On-target editing in primary T-cells via Lipid Nanoparticle (LNP) delivery
Figure 5. T-cell receptor knockout in T-cells by LNP delivery of Cas9 mRNA and synthetic TRAC-targeting sgRNA. Cas9 mRNA and synthetic TRAC sgRNA were encapsulated into lipid nanoparticles (LNP) using the GenVoy-ILM™ T Cell Kit on the NanoAssemblr™ Ignite instrument, following manufacturer’s instructions. Briefly, RNA working solutions were prepared in the supplied formulation buffer, and Cas9 mRNA or sgRNA were individually loaded into LNPs. The lipid phase and mRNA phase were combined in the Ignite instrument and downstream processed as recommended in the instructions. Primary T-cells were thawed and activated 72 hours prior to LNP addition. Cells were seeded at 0.1 x 106 cells/mL in serum-free media. Cas9 mRNA-LNPs and sgRNA-LNPS were mixed 1:1 by RNA weight and added directly to activated T-cells on Day 3. TCR knockout efficiency was assessed on Day 4 by flow cytometry after live-dead discrimination.
On-target editing in primary iPSC cells
Figure 6. On-target editing in iPS cells. Cas9 mRNA (1µg ) and sgRNA (2 µM) targeting PD1 (panel A) or TRAC (panel B) were delivered by nucleofection into iPS cells using the Lonza Nucleofection™ System. Genomic DNA (gDNA) was extracted 48 hours post nucleofection and editing was assessed by Next Generation Sequencing (NGS).
On-target editing in immortalized K562 cells
Figure 7. On-target editing in K562 cells. Cas9 mRNA (1 µg) and sgRNA(2 µM) targeting B2M (panel A) or TRAC (panel B) were delivered by nucleofection into K562 cells using the Lonza Nucleofection™ System. Genomic DNA (gDNA) was extracted 48 hours post nucleofection and editing was assessed by NGS.
Resources
Frequently asked questions
Which species-specific Cas9 is encoded by the mRNA?
Our Cas9 mRNA product encodes Cas9 from Streptococcus pyogenes (S.p.).
What is N1-Methylpseudouridine, and why is it incorporated into Cas9 mRNA?
N1-Methylpseudouridine is a modified uridine nucleoside which reduces immunogenicity while simultaneously increasing translational capacity and stability.
Does Cas9 mRNA include a 5' cap structure, poly(A) tail, untranslated regions (UTRs), and a nuclear localization signal (NLS)?
Cas9 mRNA features a Cap-1 structure at its 5’ end and includes both 5’ and 3’ untranslated regions (UTRs), a polyadenylated (PolyA) tail of at least 100 nucleotides, a nuclear localization signal (NLS), and is chemically modified with N1-Methylpseudouridine.
What delivery method is recommended for Cas9 mRNA?
The best delivery method for Cas9 mRNA depends on the application. For tissue culture experiments, we recommend electroporation. Cas9 mRNA delivery is also compatible with lipid nanoparticle (LNP) delivery.
What is the difference between enzymatic capping and CleanCap® mRNA capping?
CleanCap® is a patented mRNA capping technology developed by TriLink BioTechnologies that facilitates co-transcriptional capping of mRNA during in vitro transcription. Enzymatic capping is a post-transcriptional process during which the mRNA undergoes a series of enzymatic reactions.