Fluorescent Tag Knock-in
Precise knock-in of fluorescent tags at endogenous loci using CRISPR/Cas9 enables real-time visualization and dynamic tracking of proteins under native expression conditions.Compared to plasmid overexpression systems, Fluorescent Tag Knock-in avoids expression level distortion and mislocalization, making it suitable for high-precision imaging and quantitative analysis.Based on the FLASH-KI™ technology platform, we achieve efficient co-delivery of Cas9 RNP and Donor templates, improving knock-in efficiency and stability while maintaining ≥90% cell viability.
Service Details
| Cell Types | Wide range including tumor cell lines, normal somatic cell lines, stem cells, etc. |
|---|---|
| Services | Single-gene Fluorescent Tag knock-in (PEGFP, mCherry, mNeonGreen, TagRFP, etc.), dual-tag combinations (e.g., EGFP + FLAG) |
| Deliverables | 1 monoclonal cell line + parental cell line (2 vials/cell line, 1×10^6 cells/vial) |
| Turnaround / Price |
 Consult online for details |
Why Choose Our Knock-in Tag Service?
Core Performance Metrics
| Metric | Value |
|---|---|
| Knock-in efficiency | 45%–88% |
| Cell viability | ≥90% |
| Project success rate | 83% |
| Completed Projects | 100+ |
Service Advantages
protein behavior
Retains promoter and regulatory elements
Avoids overexpression artifacts
Avoids overexpression artifacts
HDR efficiency enhancement
With KI Enhance Drug
Peak efficiency approximately 7 days post-transfection
Peak efficiency approximately 7 days post-transfection
Efficient Co-Delivery (FLASH-KI™)
Cas9 RNP + Donor co-delivery
No electroporation/lipofection required
Significantly improved transfection efficiency
No electroporation/lipofection required
Significantly improved transfection efficiency
Low toxicity, broad compatibility
Cell viability ≥90%
Applicable to:
- Tumor cells
- Stem cells (iPSC)
- Immune cells
Tag Types and Applications
| Tags | EGFP, mCherry, mNeonGreen, TagRFP |
|---|---|
| Insertion position | N-terminal / C-terminal / internal loop |
| Applications | Live-cell imaging, protein colocalization analysis, FACS sorting |
| Features | Endogenous expression; fluorescence intensity correlates with expression level |
Workflow
Case Study
EGFP-Tag knock-in at the GAPDH locus in HEK293T cells
Goal: C-terminal EGFP knock-in at GAPDH for real-time tracking of glycolytic flux in live cells.
Approach: FLASH-KI™ delivery of Cas9 RNP + Donor vector containing EGFP and homology arms, with KI Enhance Drug.
Result: 88% knock-in efficiency in polyclonal population; stable expression clone successfully obtained.
Approach: FLASH-KI™ delivery of Cas9 RNP + Donor vector containing EGFP and homology arms, with KI Enhance Drug.
Result: 88% knock-in efficiency in polyclonal population; stable expression clone successfully obtained.
88% EGFP-KI in HEK293T cell pool by Flash-KI
Advantage and Characteristic
Optimazied Strategy
We have create a unique sgRNA Design Logic
Optimazied Strategy
We have create a unique sgRNA Design Logic
Optimazied Strategy
We have create a unique sgRNA Design Logic
Optimazied Strategy
We have create a unique sgRNA Design Logic
FAQ
Do you support multi-color labeling?
Yes; e.g., EGFP + mCherry.
Can it be used for live-cell imaging?
Yes, suitable for long-term dynamic tracking.
Does the fluorescent tag affect protein function?
- C-terminal insertion is preferred
- Linkers are used to minimize structural impact
- Control designs are available
Is it suitable for hard-to-transfect cells?
Yes; FLASH technology performs stably in sensitive cells.
