CRISPR IVT mRNA

CRISPR-mediated gene editing is a popular technique that can quickly and efficiently create mutations at target sites of a genome. VectorBuilder provides a variety of fully validated CRISPR mRNA products that ensure high in vitro expression.

Ordering Information Price Match

HiExpress™ hSpCas9

hSpCas9

HiExpress™ ABE mRNA

Our codon-optimized HiExpress™ hSpCas9 IVT mRNA ensures consistent, high expression in vitro.

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

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.

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

ABE, or adenine base editor, is a next-generation gene editing tool derived from CRISPR/Cas9 and adenine deaminase. Our codon-optimized HiExpress™ ABE IVT mRNA, based on ABE7.10, is designed to ensure high expression in vitro.

Click here to view the amino acid sequence translated from HiExpress™ ABE 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

HiExpress™ hSpCas9
hSpCas9
HiExpress™ ABE mRNA

HiExpress™ hSpCas9 IVT mRNA developed by VectorBuilder has high expression and editing efficiency_2

Figure 1. 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 1 ug of either hSpCas9 IVT mRNA, codon-optimized HiExpress™ hSpCas9 IVT mRNA, or a non-treated control (NC). (C) T7E1 editing assay in HEK293T cells 24 hours post-transfection with 1 ug 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.

Validation of hSpCas9 mRNA in vitro

Figure 2. 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 Adenine Base Editor (ABE) mRNAs in vitro

Figure 3. Validation of HiExpress™ ABE7.10 mRNA in vitro. mRNAs coding for the original and HiExpress™ ABE7.10 were co-transfected with identical gRNAs to HEK293T cells. Genomic DNA was extracted 48 hr post-transfection, and the target region was amplified by PCR and subjected to NGS. A to G editing (asterisks) was detected within the editing windows (dashed box). HiExpress™ ABE7.10 mRNA achieved a 97.7% editing rate on the 5th nucleotide, far surpassing the original’s 30.1%.