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ESpCas9 MRNA(Cap1, M1Ψ)

ESpCas9 MRNA(Cap1, M1Ψ)

eSpCas9 mRNA and TRAC sgRNA (molar ratio 1:10) were transfected into 2×105 HEK293T cells using LNP. 48h after transfection, cells were collected to extract gDNA. Genome editing efficiency were verified by PCR, Sanger sequencing and ICE analysis.

ESpCas9 MRNA(Cap1, M1Ψ)

Expression of eSpCas9 mRNA in A549 cells. The eSpCas9 expression was measured 48 hours after transfection by ELISA.

eSpCas9 mRNA(Cap1, m1Ψ)

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The eSpCas9 mRNA expresses enhanced-specificity SpCas9 protein, with sequence originally from Slaymaker et al Science. 2015 Dec 1. eSpCas9 can reduce off-target effects by over 10-fold, while maintaining robust on-target genome editing efficiency. This mRNA is capped with Cap1 structure with high capping efficiency. It has 100% substituted with N1-methyl-pseudoUridine for enhanced expression and reduced immunogenicity. The mRNA has a 100A tail in its sequence, mimics a mature mRNA.
RP-A00018
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Description

The eSpCas9 mRNA expresses enhanced-specificity SpCas9 protein, with sequence originally from Slaymaker et al., Science (2015 Dec 1). eSpCas9 significantly reduces off-target effects (by more than 10-fold), while maintaining high on-target genome editing efficiency.  
 
This mRNA features: 
- A Cap 1 structure with high capping efficiency 
- 100% substitution with N1-methyl-pseudouridine (m1Ψ) for improved protein expression and reduced innate immune response 
- GenScript’s patented UTR to enhance translation 
- A 100A poly(A) tail to mimic natural mature mRNA 
 
This construct offers high-precision genome editing with minimized off-target activity, making it ideal for CRISPR validation, therapeutic research, and functional genomics studies.
Properties
Form Liquid
Concentration 1mg/mL
Full mRNA length 4479nt
Full mRNA Molecular Weight 1457890
Storage buffer 1mM Sodium citrate, pH6.5
Storage condition Store at -20°C for short term (<3 months), store at -80°C for long term.

Quality Control Specifications
Appearance Clear and free of foreign particles
RNA Length Expected size band detected
RNA Content Target ± 5%
Integrity ≥ 75%
OD260/OD280 1.70 ~ 2.30
Capping Efficiency ≥ 90%
Endotoxin < 10 EU/mg
pH Target ± 0.5

Applications
Transfect a total of 0.5-1.5 µg of eSpCas9 mRNA and sgRNA per well (recommended molar ratio 1:10), using a transfection reagent, electroporation, or LNP encapsulation in a 24-well plate (~2 × 10⁵ cells/well). Assess genome editing efficiency 48-72 hours post-transfection via sequencing and ICE analysis.

Examples
  • ESpCas9 MRNA(Cap1, M1Ψ)
    • ESpCas9 MRNA(Cap1, M1Ψ)

  • ESpCas9 MRNA(Cap1, M1Ψ)
    • ESpCas9 MRNA(Cap1, M1Ψ)

    eSpCas9 mRNA and TRAC sgRNA (molar ratio 1:10) were transfected into 2×105 HEK293T cells using LNP. 48h after transfection, cells were collected to extract gDNA. Genome editing efficiency were verified by PCR, Sanger sequencing and ICE analysis.

  • ESpCas9 MRNA(Cap1, M1Ψ)
    • ESpCas9 MRNA(Cap1, M1Ψ)

    Expression of eSpCas9 mRNA in A549 cells. The eSpCas9 expression was measured 48 hours after transfection by ELISA.


For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.