Cas9-HA mRNA-LNP

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Description

Cas9 (CRISPR-associated protein 9) is a 160 kD protein that plays an important role in some bacteria to defend against DNA viruses and plasmids, and is heavily used in genetic engineering applications as known as “CRISPR-Cas9 genome editing”. Cas9 has a two-lobed structure, with the guide RNA positioned between the alpha-helical lobe and the nuclease lobe. The two lobes are helically connected by a single bridge. There are two nuclease domains in the multidomain nuclease lobe, the RuvC that cleaves the non-target DNA strand and the HNH nuclease domain that cleaves the target strand of DNA. CRISPR/Cas9 gene editing enables scientists to edit genomic DNA sequences, adding, deleting or altering them. This product is designed as a tool for the delivery and expression of HA-tagged CAS9 mRNA for research. The product leverages the lipid nanoparticle (LNP) technology platform for simple and efficient delivery of HA-tagged CAS9 mRNA to a variety of mammalian cells in vitro and in vivo. The LNPs used are formulated with SM-102, DSPC, cholesterol and DMG-PEG2000 at an optimal molar concentration for a high rate of encapsulation and efficient mRNA delivery. The sequence of the CAR9-HA protein consists of CAR9 containing 1368 amino acids and a C-terminal HA tag (YPYDVPDYA), and the GenPept accession number for CAR9 is WP_038431314.

Product Overview

Composition

mRNA-LNPs suspended in PBS (-Ca, -Mg) (pH: 7.0-7.4).

Cell Type Tested

HEK293S, T cells, iPSC

Application & Handing

Upon receiving product, briefly pulse spin before opening to ensure product is at bottom of container. It is important not to spin for too long as this may rupture mRNA-LNPs. Do not vortex. Work with mRNA-LNPs on ice and minimize the time that the product spends at room temperature. After handling the product during experiments, return immediately to ice. mRNA-LNP products should only be handled with certified RNase-free reagents and consumables. Use of filtered pipette tips is highly recommended.

Storage

4°C; ice

Product Image

CAS9 Protein repairs DNA by Knockout

Figure 1: Pathway on the left shows Knock-out of genes by NHEJ. Pathway on the right shows knock-in of genes.

FACS Analysis for CAS9-mRNA-LNP

Figure 2: Shows TRAC gene expression measured by FACS in T cells with various amounts of Cas9-mRNA-LNP. Six days after electroporation, the T cells treated with 4x the usual volume of Trilink Cas9 RNA LNPs had 80% as much TRAC knockout as cells treated with Cas9 protein (62%- 75%). “

For Research Use Only. Not for use in diagnostic procedures.