hSpCas9 IVT mRNA

Name: hSpCas9 IVT mRNA
Brand: VectorBuilder

VectorBuilder's hSpCas9 IVT mRNA can be ordered with or without the modified nucleotide, N1-Methylpseudouridine (m1Ψ) and its gene editing function has been functionally validated in cell culture.

Ordering Information Price Match

Click here to view the amino acid sequence translated from hSpCas9 mRNA >>

Shipping and storage

Our IVT mRNA products are stored in a 1 mM sodium citrate buffer (pH 6.4) and can be stored at -80°C for up to 12 months. RNA products are shipped on dry ice and freeze-thaw cycles should be avoided.

Experimental Validation

Co-transfected hSpCas9 mRNA and gRNA to 293T-EGFP cells and analyzed EGFP expression level by microscopy and flow cytometry.

Figure 1. Validation of hSpCas9 mRNA in vitro. (A) IVT Cas9 mRNA was transfected into 293T-EGFP cells with two types of EGFP-targeting gRNA. EGFP expression in non-treated (NC) and transfected cells was observed by microscopy (B) and quantified using flow cytometry (C + D). (E + F) The editing to EGFP genes on the genome was further confirmed by T7E1 assay and Sanger sequencing.

Validation_of_hSpCas9_mRNA_knockout_with_dual_gRNAs

Figure 2. HiExpress™ hSpCas9 IVT mRNA developed by VectorBuilder has high expression and editing efficiency. (A) Western blot analysis and (B) quantification of normalized Cas9 expression relative to hSpCas9 in HEK293T cells 24 hours post-transfection with 1000 ng of either hSpCas9 IVT mRNA, codon-optimized HiExpress™ hSpCas9 IVT mRNA, or a non-treated control (NC). (C) T7E1 editing assay (D) and quantification of editing efficiency (%) in HEK293T cells 24 hours post-transfection with 1000 ng of gRNA and the indicated amounts of hSpCas9 or HiExpress™ hSpCas9 IVT mRNA. The blue star indicates the intact PCR amplicon and the pink arrows indicate the fragments generated by T7E1.