Professional technology integration to support the entire R&D process

Gene knock-in is a cutting-edge genetic engineering technique that involves the precise insertion of a new gene or DNA sequence into a specific, predetermined location within the genome. This powerful method enables researchers to study gene function, create accurate disease models, and develop advanced therapeutic strategies. Utilizing technologies like CRISPR/Cas9, homologous recombination, and viral vectors, our service provides efficient and reliable targeted integration for your experimental or therapeutic goals.

The process is driven by the revolutionary CRISPR-Cas9 system. A designed single-guide RNA (sgRNA) directs the Cas9 endonuclease to create a precise double-strand break (DSB) at the target genomic locus. The cell's natural repair machinery is then activated. When a donor DNA template with homologous arms is provided, the cell uses the Homology-Directed Repair (HDR) pathway to copy this template while repairing the break. This allows for the exact integration of your gene of interest or specific mutation into the genome.

We offer a complete, end-to-end solution to ensure successful gene integration and cell line development.

Knock-In Vector Design & Construction: Our experts design the optimal vector (plasmid, viral, or CRISPR-compatible) carrying your target sequence. We optimize homology arm length, select efficient integration sites, and include necessary elements (e.g., reporters, selectable markers) for high specificity and success.

Cell Transfection/Delivery: We introduce the engineered knock-in construct into your target cells using optimized methods such as electroporation, chemical transfection, or viral transduction (AAV, Lentivirus). Our protocols are tailored to maximize efficiency while preserving cell viability.

Screening of Positive Cells: Post-delivery, we screen the cell population to identify clones with successful gene integration. Screening is performed using robust methods like antibiotic selection, fluorescence-activated cell sorting (FACS), or PCR-based genotyping.

Monoclonal Cell Line Generation: To ensure genetic uniformity and stability, we perform single-cell cloning via limiting dilution or FACS-based sorting. This step is critical for establishing pure, clonal cell lines.

Comprehensive Validation & Delivery: Each monoclonal cell line undergoes rigorous verification. We confirm precise genomic insertion, sequence accuracy, and functional protein expression using PCR, Sanger sequencing, qPCR, Western blot, and other relevant functional assays. Finally, we deliver the fully validated, ready-to-use cell line along with a detailed report.

Gene Function Research
Knock-in cell lines can be used to study the function and mechanism of action of specific genes in
cellular processes, revealing the biological function of genes through overexpression or modification.

Cell Signaling Pathway Research
Knock-in cell lines can be used to study the regulatory mechanisms and interactions of cell signaling pathways, thereby gaining a deeper understanding of cell biological processes and the mechanisms of disease development.

Disease Establishment Model
By establishing cell lines that mimic disease with specific gene knock-in, it is possible to study
disease pathogenesis, screen therapeutic targets, and develop new drugs.

Drug Target Identification
Knock-in cell lines can be used to evaluate the effects of drug targets and drug efficacy, providing
important reference and support for drug development.

Our Gene Knock-In Service provides a reliable and efficient pathway to achieve precise genomic integrations. From initial vector design to final validation of monoclonal cell lines, we combine advanced CRISPR-Cas9 technology with optimized protocols to deliver high-quality, reproducible results. Whether for functional genomics, disease modeling, or cell therapy development, our service accelerates your research with accuracy and expertise.

If you are interested in ordering, please contact us
Email: sales@reqbio.com