Flash-KI Platform

Technology Platform Overview

High-Efficiency Targeted Integration Platform (FLASH-KI): Precise Writing, Unprecedented Efficiency

Gene knock-in (KI) is a critical technology for endogenous gene fluorescent tagging, reporter gene insertion, and site-directed mutagenesis. It is widely used in dynamic protein tracking, cell line development, disease modeling, and drug screening. However, conventional KI methods rely on Homology-Directed Repair (HDR), which notoriously suffers from extremely low efficiency in most cell types, high cytotoxicity, and prolonged screening cycles—making it the primary bottleneck in cell model generation.
To address these pain points, EDITGENE has developed the FLASH-KI technology platform, integrating two core technologies: a high-efficiency delivery system and cellular repair pathway modulation. By combining precision editing components with transient efficiency-enhancing strategies, FLASH-KI achieves highly efficient, low-toxicity, and selection-free precise knock-in across a variety of hard-to-transfect cells (e.g., hESCs, iPSCs) and complex genomic loci.
Service Advantages

Core Technology 1: FLASH-KI Delivery System

Conventional knock-in strategies typically require separate delivery of Cas9 protein, sgRNA, and Donor templates, resulting in cumbersome procedures and limited efficiency. EDITGENE’s proprietary FLASH-KI delivery vector enables the simultaneous delivery of the RNP complex and Donor DNA, significantly streamlining the workflow and boosting synergistic efficiency.

Technical Advantages
•Co-delivery Design: Synchronized entry of RNPs and Donor DNA into the cell enhances the spatiotemporal matching of the repair template with the editing site.
•Low Cytotoxicity: The protein-based delivery approach avoids immunogenicity and random integration risks associated with DNA plasmids.
•Broad Applicability: Validated in over 400 cell lines, including hard-to-transfect types like iPSCs, hESCs, and organoids.
•Drug Selection-Free: Achieves high-purity cell pools without the need for enrichment across multiple validated targets.

Core Technology 2: NHEJ Inhibitor (KI Enhancer Drug)

During CRISPR-mediated DNA double-strand break repair, Non-Homologous End Joining (NHEJ) is the dominant repair pathway, while HDR efficiency is typically below 10%. EDITGENE has independently developed an NHEJ inhibitor that transiently and reversibly suppresses key NHEJ proteins during the editing process, forcing the cell to prioritize the HDR pathway.

Technical Advantages
•Significant Efficiency Boost: Validated across multiple cell lines, increasing KI efficiency by 5-to-7-fold.
•Reversible Inhibition: The drug's duration of action is controllable, ensuring no impact on long-term cell viability or function.
•No Genetic Modification Required: Independent of prior cell line modifications, making it universally applicable to any cell type.
•High Compatibility: Can be seamlessly integrated with FLASH-KI, electroporation, lipofection, and other delivery methods.

Service Advantages:

FLASH-KI vs. Conventional Methods

Dimension Key Scientific Question Our Technology Key Resources
Point Mutations Which variants are true drivers of tumorigenesis? Precision genome editing Mutation cell model library
Tumor Dependency Which genes are essential for cancer cell survival? Gene dependency profiling Tumor dependency evaluation platform
Drug Targets Which genes are viable therapeutic targets? Target validation, functional assessment Knockout cell line panels
Note: Conventional HDR data is based on industry literature and internal validation; conventional electroporation data reflects empirical results from select cell lines; FLASH-KI data is derived from EDITGENE internal testing.

Expected Performance Across Different Cell Types (FLASH-KI Platform)

Difficulty Level Representative Cell Lines Expected KI Efficiency (Pool) Project Success Rate (Homozygous Clones) Turnaround Time Key Influencing Factors
⭐Low
(Standard Immortalized)		 HEK293, HeLa, CHO, HCT-116 60-90% >90% 8-10 weeks Target locus, Donor design quality
⭐⭐Medium
(Hard-to-transfect, some suspension)		 A9, THP-1, K562, Jurkat 30-60% 70-80% 10-12 weeks Cell state, transfection parameters, NHEJ inhibitor concentration
⭐⭐⭐High
(Stem cells, Primary, Organoids)		 iPSC, hESC, Primary T cells, Organoids 15-30% 60-70% 12-14 weeks Cell health, monoclonal formation efficiency
Definitions:
• Pool Efficiency: The proportion of cells with correct knock-in within the unselected post-transfection population (assessed via flow cytometry or sequencing).
• Project Success Rate: The percentage of projects that successfully deliver at least one homozygous monoclonal cell line.
• Data is based on statistics from 100+ completed KI projects by EDITGENE.
Service Types
Standard KI: Monoclonal cell line + Sequencing validation.

High-Purity Cell Pool: For easily transfected cells and housekeeping genes (e.g., GAPDH, ACTB), we directly deliver a cell pool with >80% positivity, saving time on monoclonal screening.

Custom Hard-to-Transfect Cells: Exclusive optimized protocols for hESCs, iPSCs, and primary cells to ensure the generation of clean, single-background positive monoclonal cell

Technical Workflow

Application Cases
  1. As of Q1 2026, the FLASH-KI platform has successfully delivered 20+ commercial projects covering immortalized cells, tumor cells, and stem cells, achieving precise integrations of fragments ranging from 0.5 to 5.5 kb. The overall project success rate is 98.6%, with a 100% success rate for fragments under 5500 bp.

Case 1: Precise KI of a High-Abundance Gene (HEK293T-GAPDH-EGFP)

Client Need: Insert an EGFP fluorescent tag (841 bp) at the C-terminus of the GAPDH gene in HEK293T cells for live-cell dynamic tracking, requiring a high-purity positive pool without drug selection.

Technical Breakthrough: Conventional electroporation yielded only 68% positivity with 30% cell death. FLASH-KI achieved: ① 88% positivity without antibiotic screening; ② >95% cell viability; ③ 40% improvement in fluorescence signal uniformity.

Timeline: Delivered an 88% positive polyclonal pool in 4 weeks; obtained homozygous KI monoclonal cell lines in 8 weeks (30% faster than the industry average).


Case 2: Complete Gene KI at a Random Target (A-9-T1R1-CoGFP-Puro)

Client Need: Insert an EF1a-CoGFP-T2A-Puro cassette (2115 bp) into the T1R1 locus of A-9 cells while preserving native receptor conformation and signaling pathway integrity.

Technical Breakthrough: FLASH-KI achieved: ① 46% positivity without selection markers; ② >95% cell viability; ③ Monoclonal stability exceeding 15 passages.

Timeline: Functional polyclonal pool in 5 weeks; monoclonal screening and validation completed in 9 weeks.

Case 3: Human Embryonic Stem Cell Editing (hESC-H9-TH-P2A-EGFP)

Client Need: Insert a P2A-NeoR-EF1a-EGFP cassette (2283 bp) at the C-terminus of the TH gene in hESC-H9 cells, maintaining pluripotency and differentiation potential.

Technical Breakthrough: Overcame barriers for hard-to-transfect stem cells: ① 20% positivity without drug selection (industry average <5%); ② >90% cell viability; ③ Normal expression of Oct4/Sox2 pluripotency markers.

Timeline: 20% positive polyclonal pool in 5 weeks; monoclonal identification completed in 11 weeks.

CRISPR-iSCREEN™ Library in Stock
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Library Plasmid

coverage rate > 99%, uniformity < 10

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Library Plasmid

coverage rate > 99%, uniformity < 10

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Library Plasmid

coverage rate > 99%, uniformity < 10

$99
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