hSpCas9 IVT mRNA

hSpCas9 mRNA was generated with or without modified nucleotide, N1-Methylpseudouridine (m1Ψ). Its gene editing function has been fully validated in cell culture.

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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. The products are shipped on dry ice and should be avoided for freeze-thaw cycles.

Experimental Validation

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

In vitro gene editing using Cas9 mRNA

(A) IVT Cas9 mRNA was generated with modified nucleotides, N1-Methylpseudouridine (m1Ψ), and 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 (100X) (B) and quantified using flow cytometry (C). (D) At 48 or 72 h post transfection, the EGFP expression was reduced to about 40% of the non-treated 293T-EGFP cells. (E and F) The editing to EGFP genes on the genome was further confirmed by T7E1 assay (E) and Sanger sequencing (F). The highlighted adenine indicates the gene editing introduced an insertion in the sequence.

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