Getting started with CRISPR? Need proven CRISPR validated protocols? Our CRISPR Validated Protocols collection has been developed by our expert R&D team. We have matched each of the validated and optimized protocols with the best products to help reduce trial and error and to accelerate your rate of discovery. These step-by-step CRISPR protocols have been optimized for maximum efficiency, viability, and reproducibility across a broad range of cell types and gene targets. We truly want you to succeed with your CRISPR experiment the first time and have confidence in your data.
TrueCut Cas9 Protein v2
Next generation Cas9 protein designed to deliver maximum editing efficiency
Introducing Invitrogen TrueCut Cas9 Protein v2, a wild type Cas9 in protein form designed to deliver consistently higher editing efficiency across a range of gene targets and cell types. Based on 30 years of industry-leading innovations, our investment in R&D helps ensure that our products reliably fulfill your research needs:
- Consistently high editing efficiency in all tested cell lines including standard, immune, primary, and stem.
- Up to 2X higher editing efficiency in difficult targets than the competition
- High quality—manufactured under strict ISO 13485 quality standards
- Validated protocols—achieve success faster using protocols optimized for an extensive list of cell types
Truly stunning performance, reliable results, and more choices—your research demands nothing less
Achieve functional knockout in up to 90% of transfected human primary T-cells
Figure 1. T cell receptor knockout using TrueGuide synthetic gRNAs. Human T cells were isolated and activated using Invitrogen Dynabeads magnetic beads (Cat. Nos. 11344D and 11141D, respectively) and then transfected with TrueCut Cas9 Protein v2 and TrueGuide Synthetic gRNA for T cell receptor alpha (TRAC) or beta (TRBC) regions using the Invitrogen Neon Transfection System. Following transfection, editing efficiency was measured by the Invitrogen GeneArt Cleavage Detection assay (A), or by measuring % T cell receptor negative (TCR–) cells using the Invitrogen Attune NxT Flow Cytometer (B). Cells analyzed by flow cytometry were stained with a T cell receptor–specific antibody conjugated to PE (C).
Consistently outperforms the competition
Figure 2. Invitrogen CRISPR genome-editing tools consistently outperform products from other suppliers. Genome editing of multiple gene targets was performed with TrueCut Cas9 Protein v2 and corresponding TrueGuide Synthetic gRNAs. Delivery was achieved using optimized transfection protocols and Invitrogen Lipofectamine CRISPRMAX Transfection Reagent in two cell lines, (A) A549, a human lung carcinoma cell line and (B) MDA-MB231, a human breast cancer cell line. The graphs compare the same experiments using products and recommended protocols from another manufacturer. With the Invitrogen system, cleavage efficiency is improved for low-efficient loci (PRKCG-T1 and CMPK1-T1) and shows consistently greater efficiency when compared to products and protocols from other suppliers, even for challenging loci.
Robust editing efficiencies in cancer cell lines
Figure 3. Robust editing efficiencies in cancer cells. A wide range of cancer cell lines and gene targets were tested to determine editing efficiencies using the modified single TrueGuide Synthetic gRNA (Cat. No. A35511) complexed with TrueCut Cas9 Protein v2 (Cat. No. A36497). Here, the Cas9 RNP complex was delivered into three different cancer cell lines using Lipofectamine CRISPRMAX Cas9 Transfection Reagent (Cat. No. CMAX00003) and, at 72 hours posttransfection, cells were harvested and tested for cleavage efficiency using either the GeneArt Genomic Cleavage Dectection Assay (GCD) or Ion Torrent Next-Generation Sequencing (NGS). Panel ( A) A549, an epithelial lung carcinoma cells, (B) U2OS, osteosarcoma line, (C) MDA-MB231, epithelial (breast) adenocarcinoma line
Reliable editing efficiencies across a broad range of cell lines and with different delivery methods
Figure 4. Consistent editing efficiency across a wide range of cell types. TrueCut Cas9 Protein v2 has been tested using lipid-mediated delivery or the Neon electroporation transfection system across a wide range of cell types. Here we demonstrate high efficiency editing in four representative cell types including adherent cell lines (A549, HepG2), suspension cells (THP1) and primary immune cells (Primary Human T Cells). Cell lines were transfected with TrueCut Cas9 Protein v2 (Cat. No.A36497) and the TrueGuide Positive Control crRNA (Cat. No.A35517) and TrueGuide tracrRNA (Cat. No. A35506).
Our R&D scientists demonstrated that by combining Cas9 protein with CRISPR gRNAs and complexing them with a lipid-based transfection reagent, you can significantly simplify the cell engineering workflow and achieve cleavage efficiencies of up to 85% in many cell lines (Liang et al. 2015).
The Cas9 RNP complex can act immediately after it enters the cell, since transcription and translation are not required. Moreover, the complex is rapidly cleared from the cell, minimizing the chance for off-target cleavage events when compared to vector-based systems.
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For Research Use Only. Not for use in diagnostic procedures.