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Cas9 SmartNuclease™ mRNA, Injection- and Transfection-ready

Move in vivo genome editing projects forward faster with injection- and transfection-ready Cas9 SmartNuclease mRNA

  • Ready to use for in vivo genome editing applications
  • Functionally validated Cas9 SmartNuclease
  • Backed by expert, easy-to-reach technical support
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Description
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CAS500A-1
Transfection-ready Cas9 SmartNuclease mRNA (Eukaryotic version)
20 µg
$ 364

Overview

Accelerating in vivo genome editing

From basic biological studies to generating novel disease models, SBI’s injection- and transfection-ready PrecisonX™ Cas9 SmartNuclease™ mRNA is here to move your in vivo genome engineering projects forward faster.

Use with an appropriate gRNA and, if your application needs it, a homologous recombination (HR) targeting vector for gene knock-ins, knock-outs, editing, and tagging. The Cas9 SmartNuclease will generate a double-strand break (DSB) at the site specified by the gRNA, and this DSB can be efficiently and precisely repaired in a manner that includes your desired mutation through the use of an HR targeting vector.

As with all of our Cas9 delivery options, the Cas9 SmartNuclease mRNA is functionally validated and comes backed by our expert technical support team—if you’ve got a genome engineering question just ask by emailing tech@systembio.com.

  • Ready to use for in vivo genome editing applications
  • Functionally validated Cas9 SmartNuclease
  • Backed by expert, easy-to-reach technical support

Why an HR targeting vector is a recommended

Even though gene knock-outs can result from DSBs caused by Cas9 alone, SBI recommends the use of HR targeting vectors (also called HR donor vectors) for more efficient and precise mutation. HR donors can supply elements for positive or negative selection ensuring easier identification of successful mutation events. In addition, HR donors can include up to 6-8 kb of open reading frame for gene knock-ins or tagging, and, when small mutations are included in either 5’ or 3’ homology arms, can make specific, targeted gene edits.

Efficiently generate gRNA with the T7 gRNA SmartNuclease Synthesis Kit

Pair your Cas9 SmartNuclease mRNA with our T7 gRNA SmartNuclease Synthesis Kit, which comes with a pre-linearized, ready-for-cloning gRNA expression vector and all the reagents you need for T7-driven in vitro transcription of your cloned gRNA.

Not sure whether you need a CRISPR/Cas9 plasmid, purified protein, or mRNA?

Use this table to choose the CRISPR/Cas9 product that’s right for you:

For This Application
In these types of cells

Use These Products

MODIFYING ORGANISMS

  • Gene tagging
  • Transgenic organism generation
  • Model organism engineering
Embryos—to create transgenic animals Injectable Cas9 mRNA & gRNA Synthesis Kits
Cas9 Protein
EGFP-labeled Cas9 Protein
Animals models—in vivo genome editing AAV-Cas9 Vectors  
Cas9 Protein
EGFP-labeled Cas9 Protein
MODIFYING CELL LINES

  • Stable KO, KI, and genome editing of
    somatic cells
  • Transgenic cell line generation
  • Cell-based disease models
Cells that are transfectable Cas9 Plasmids
Cas9 Protein
EGFP-labeled Cas9 Protein
Difficult-to-transfect cell lines:

  • Primary cells
  • Hematopoietic cells
  • Stem cells
AAV-Cas9 Vectors 
Lenti Cas9 Systems
SCREENING

  • Genome-wide surveys
  • gRNA library screens
  • Functional screens
All cell types requiring stable Cas9 overexpression Lenti Cas9 Systems
AAVS1 Safe Harbor Site Cas9
Gene Knock-in System
Cas9 Protein
EGFP-labeled Cas9 Protein
PRE-CLINICAL APPLICATIONS

  • Off-target events are of highest concern
All cell types and applications Cas9 Nickase, available in all delivery formats
Cas9 Protein
EGFP-labeled Cas9 Protein
SIMULTANEOUS ENGINEERING OF MULTIPLE MUTATIONS All cell types and applications Multiplex gRNA cloning kit, compatible with all Cas9 delivery options

How It Works

Genome engineering with CRISPR/Cas9

For general guidance on using CRISPR/Cas9 technology for genome engineering, take a look at our CRISPR/Cas9 tutorials as well as the following application notes:

CRISPR/Cas9 Gene Knock-Out Application Note (PDF) »
CRISPR/Cas9 Gene Editing Application Note (PDF) »
CRISPR/Cas9 Gene Tagging Application Note (PDF) »

CRISPR/Cas9 Basics

Through careful selection of the target sequence and design of a donor plasmid for homologous
recombination, you can achieve efficient and highly targeted genomic modification with CRISPR/Cas9.

The system

Cas9 protein—uses guide RNA (gRNA) to direct site-specific, double-strand DNA cleavage adjacent to a protospacer adapter motif (PAM) in the target DNA.

gRNA—RNA sequence that guides Cas9 to cleave a homologous region in the target genome. Efficient cleavage only where the gRNA homology is adjacent to a PAM.

PAM—protospacer adapter motif, NGG, is a target DNA sequence that spCas9 will cut upstream from if directed to by the gRNA.

The workflow at-a-glance

DESIGN: Select gRNA and HR donor plasmids. Choice of gRNA site and design of donor
plasmid determines whether the homologous recombination event results in a knock-out,
knock-in, edit, or tagging.

CONSTRUCT: Clone gRNA into all-in-one Cas9 vector. Clone 5’ and 3’ homology arms into HR
donor plasmid. If creating a knock-in, clone desired gene into HR donor.

CO-TRANSFECT or CO-INJECT: Introduce Cas9, gRNA, and HR Donors into the target cells
using co-transfection for plasmids, co-transduction for lentivirus, or co-injection for mRNAs.

SELECT/SCREEN: Select or screen for mutants and verify.

VALIDATE: Genotype or sequence putative mutants to verify single or biallelic conversion.

Supporting Data

Knock-in of GFP at the AAVS1 Safe Harbor Site using Cas9 SmartNuclease mRNA

Figure 1. Cas9 SmartNuclease mRNA-mediated knock-in of GFP to the AAVS1 Safe Harbor Site. METHODS: The AAVS1 gRNA sequence was cloned into the T7 gRNA Cloning and Production Vector and in vitro transcribed. These gRNAs were co-transfected with the Cas9 synthetic mRNA in combination with an AAVS1 HR Targeting Vector harboring a GFP marker. The activities of the Cas9 mRNA + AAVS1 gRNA transfection was compared with that of the EF1 Cas9 SmartNuclease-AAVS1 gRNA All-in-one vector system. Cells were imaged for GFP fluorescence after 3 days.

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