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ProteanFect™ CRISPR Gene Editing Transfection Kit
ProteanFect™ Transfection Kit offers a non-viral, non-electroporation, and non-liposomal transfection system utilizing engineered mammalian proteins. This innovative design achieves high transfection efficiency while maintaining a superior safety profile. The ProteanFect™ CRISPR-Cas9 mRNA Gene Editing Kit is specifically designed for effective gene editing by delivering Cas9 mRNA and sgRNA into difficult-to-transfect cell lines. Additionally, it excels in co-transfecting sgRNAs targeting different genes, enabling simultaneous knockout of multiple genes.
| Number | Standard | Price(USD) | Stock |
|---|---|---|---|
| PT04-0010B | 0.1 mL | 971 | In 4 weeks |
| PT04-0020B | 0.2 mL | 2092 | In 4 weeks |
| PT04-0050B | 0.5 mL | 2839 | In 4 weeks |
| PT04-0100B | 1 mL | 4444 | In 4 weeks |
| PT04-0500B | 5 mL | 19541 | In 4 weeks |
Transfection Performance
Publication
Zhu, P. et al. Efficient Generation of CAR-T By the First Coacervate-Based Gene Delivery System. ASH Annual Meeting & Exposition, (2024).
Gao, X. et al. The First Coacervate-based Delivery System for Advanced Gene Therapy. ASGCT 28th Annual Meeting Abstracts, (2025).
Gao, X. et al. The First Coacervate-Based Delivery System for CAR-T Cell Engineering and Manufacturing. Presented at ASGCT 28th Annual Meeting Abstracts.
Documents
Product Information
Components & Storage
The kit is shipped on dry ice. Once received, store the components as indicated below. The kit includes positive control samples with EGFP-encoding mRNA to verify transfection efficiency as well as single-guide RNA (sgRNA) targeting the human TRAC gene.
| Components | Storage |
|---|---|
| Reagent A (for CRISPR-Cas9 mRNA) | 2-8℃ |
| Reagent B (for CRISPR-Cas9 mRNA) | -20°C(Avoid repeated freeze-thaw cycles |
| EGFP mRNA (1 µg/µL) | -80°C(Avoid repeated freeze-thaw cycles) |
| Human TRAC -sgRNA (1 µg / µL) | -80°C (Avoid repeated freeze-thaw cycles) |
Shipping Information
Shipped on Dry Ice
Compatible Cell Lines
Cell Lines Successfully Edited Using ProteanFect™ CRISPR Gene Editing Transfection Kit
| Cell lines | Tested Nucleic Acid Types for Transfection |
|---|---|
| Jurkat T (human T lymphoblastic leukemia cells) | sgRNA |
| LX-2 (human hepatic stellate cells) | sgRNA |
| HepG2 (human liver tumor cells) | sgRNA |
| THP-1 (human acute monocytic leukemia cell line) | sgRNA |
| Raji (human Burkitt lymphoma cells) | sgRNA |
| K562 (human chronic myeloid leukemia cells) | sgRNA |
| MOLT-16 (human T lymphoblastic leukemia cells) | sgRNA |
| SH-SY5Y (human neuroblastoma cells) | sgRNA |
| U2OS (human osteosarcoma cells) | sgRNA |
| U937 (human lymphoma cell line) | sgRNA |
| HFF (human foreskin fibroblasts) | sgRNA |
| HEK-293 (human embryonic kidney cell line) | sgRNA |
| MC38 (mouse colon cancer cells) | sgRNA |
| RAW264.7 (mouse mononuclear macrophage leukemia cells) | sgRNA |
| LLC (mouse Lewis lung cancer cells) | sgRNA |
| C2C12 (mouse myoblasts) | sgRNA |
| COS7 (African green monkey kidney fibroblast-like cells) | sgRNA |
Product Data
Transfection Protocol for mRNA per Well of a 96-Well Plate
| Scheme | Step | Transfection Protocol for ProteanFect™ CRISPR Gene Editing in Various Cell Lines per Well of a 96-Well Plate |
|---|---|---|
 |
1. Transfection Complex Preparation |
1.1 Mix Reagent A (for CRISPR-Cas9 mRNA) with mRNA Mix 0.25 µg Cas9 mRNA and 0.25 µg sgRNA with 40 µL of Reagent A (for CRISPR-Cas9 mRNA). Note: Invert the Reagent A (for CRISPR-Cas9 mRNA) briefly before use to ensure uniformity. 1.2 Add Reagent B (for CRISPR-Cas9 mRNA) Add 1.4 µL of Reagent B (for CRISPR-Cas9 mRNA) to the mixture. Mix gently by pipetting up and down 20-30 times or vortexing for 10 seconds. |
 |
2. Cell Preparation |
2.1 Suspension cells Harvest the cells by centrifugation at 300 g for 5 minutes. Discard the supernatant and wash cells once with Opti-MEM. Resuspend cells with Opti-MEM and adjust concentration to 5×10⁶ - 1×10⁷ cells/mL. Note: Avoid including FBS in the transfection medium. 2.2 Adherent cells Maintain 50%-80% cell confluence. Remove medium, wash cells once with Opti-MEM, then add 20 µL of Opti-MEM. Note: Avoid including FBS in the transfection medium. Optional: Harvest cells by trypsinization, then resuspend them in Opti-MEM at a concentration of 5×10⁶ - 1×10⁷ cells/mL for subsequent transfection. |
 |
3. Transfection |
3.1 Mix transfection complex with cells For suspension cells, mix 40 µL of transfection complex with 20 µL of cell suspension and gently pipet up and down 2-3 times. For adherent cells, apply directly to the cells. 3.2 Incubation Incubate the cells with the transfection complex for 45-60 minutes in a cell culture incubator. 3.3 Termination Terminate the reaction by adding ≥200 µL of culture medium (10X cell suspension), centrifuge at 300 g for 5 minutes, and discard the supernatant. For adherent cells, replace the transfection mixture with ≥200 µL of culture medium (10X cell suspension). Note: The cell pellet may adhere to the tube walls. Gently remove the supernatant to minimize cell loss. 3.4 Post-transfection culture Incubate the transfected cells in culture medium and evaluate the editing efficiency of the target genes after 48 to 72 hours, or at an appropriate time. |
| Scheme | Step |
|---|
Technical FAQ
1.1 Optimize Transfection Parameters
Optimize transfection parameters for each cell type. Extend incubation time: Adjust the incubation time of the transfection complex with cells. The maximum incubation time is 2 hours for cell lines. Increase ProteanFect™ transfection complex: Consider increasing the amount of transfection complex to improve transfection efficiency.
1.2 Improve Cell Condition
For cell lines, transfect cells with >90% viability, confirmed by trypan blue exclusion. Avoid using cells beyond 15 passages, and allow 2-3 passages for recently thawed cells to stabilize before transfection.
1.3 Use Positive Control
We recommend using a 96-well plate format to optimize transfection conditions for a specific cell type, with EGFP mRNA as the positive control.
Optimize transfection parameters for each cell type. Extend incubation time: Adjust the incubation time of the transfection complex with cells. The maximum incubation time is 2 hours for cell lines. Increase ProteanFect™ transfection complex: Consider increasing the amount of transfection complex to improve transfection efficiency.
1.2 Improve Cell Condition
For cell lines, transfect cells with >90% viability, confirmed by trypan blue exclusion. Avoid using cells beyond 15 passages, and allow 2-3 passages for recently thawed cells to stabilize before transfection.
1.3 Use Positive Control
We recommend using a 96-well plate format to optimize transfection conditions for a specific cell type, with EGFP mRNA as the positive control.
Transfected cells may exhibit transient changes in behavior, but typically, viability should be restored by the second day post-transfection.
In 96-well formats, it is common for the pellet to be less distinct and may adhere to the tube walls. Gently pipetting can help minimize cell loss.
For further questions, please contact us at: tech@nanoportalbio.com.