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Exogenous Gene Integration Site Analysis Service
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Exogenous Gene Integration Site Analysis Service

With the rapid advancement of genetic engineering technology, the introduction and integration of exogenous genes has become a key tool in biomedical research, gene therapy, and agricultural breeding. However, the integration site of an exogenous gene within the host genome has a crucial impact on its expression stability, functional performance, and safety. Therefore, exogenous gene Integration Site Analysis services have emerged as a crucial tool for ensuring the safety and effectiveness of genetic manipulation.technique.

Definition

Exogenous gene Integration Site Analysis uses molecular biology techniques to precisely identify the insertion location, copy number, and integration status of an exogenous gene within the host cell genome. This test not only examines whether the exogenous gene has successfully integrated, but also focuses on analyzing the characteristics of the integration site, such as whether it is located in the gene coding region, regulatory region, or repetitive sequence, thereby assessing its potential impact on host gene function.

Detection Principle

The core principles of exogenous gene Integration Site Analysis are based on nucleic acid hybridization, PCR amplification, and high-throughput sequencing technologies. Currently, mainstream methods include:

1. PCR-based methods, such as inverse PCR and ligation-mediated PCR, use specific primers to amplify the junction region between the exogenous gene and the host genome, thereby identifying the integration site.

2. High-throughput sequencing technologies, such as whole-genome sequencing (WGS), targeted sequencing, and chromatin conformation capture (Hi-C), enable high-throughput and high-precision identification of exogenous gene integration sites across the entire genome, while simultaneously analyzing copy number and structural variation.

3. Fluorescence in situ hybridization (FISH): Fluorescently labeled probes hybridize to host chromosomes to directly visualize the physical location of the exogenous gene.

The combined application of these technologies enables comprehensive and accurate analysis of the integration status of the exogenous gene.
 
 

Workflow

A complete exogenous gene Integration Site Analysis service typically includes the following steps:
  • 1.Sample preparation
    Extract genomic DNA from the cells to be tested, ensuring that the DNA quality meets the test requirements.
  • 2.Library construction
    Construct a sequencing or PCR library, depending on the test method. For example, in WGS, genomic DNA needs to be fragmented and sequencing adapters added.
  • 3. Sequencing or Amplification
    Whole-genome sequencing is performed using a high-throughput instrument, or the target region is amplified using PCR.
  • 4.Data Analysis
    Bioinformatics tools are used to align sequencing data, identify the boundary sequences between the exogenous gene and the host genome, determine the integration site, and assess copy number, insertion orientation, and nearby gene function.
  • 5.Result Verification
    Key integration sites are verified using techniques such as Sanger sequencing, qPCR, or FISH to ensure the reliability of the results.
  • 6.Report Generation
    A detailed test report is provided, including integration site coordinates, genomic annotations, potential risk analysis, and visual charts.
Service Advantages

High Precision and High Throughput

Leveraging next-generation sequencing technology, multiple samples can be analyzed simultaneously, accurately identifying integration sites across the entire genome, even detecting low-frequency integration events.

Comprehensiveness

In addition to providing integration site information, it also analyzes copy number, insertion structure, and potential effects on host genes (such as whether it disrupts tumor suppressor genes or activates oncogenes).

Efficiency and Automation

Leveraging proven bioinformatics pipelines, it rapidly processes large amounts of data, shortens analysis cycles, and improves research efficiency.

Safety Assessment

Integration site annotation enables prediction of potential risks associated with exogenous gene insertion, providing a key basis for safety assessments of genetic manipulation.

Customized Services

Targeted solutions can be provided based on customer needs, such as optimization for specific cell types and multi-omics integrated analysis.

Application Scenarios

Exogenous gene Integration Site Analysis services have a wide range of applications in various fields:
Service-Application
 Gene Therapy: In CAR-T cell therapy and viral vector-mediated gene therapy, detecting the integration site of the exogenous gene helps assess the risk of insertional mutagenesis and ensure treatment safety.
 Transgenic Organism Research: In agricultural transgenic crop and livestock breeding, integration site analysis can optimize transgene design, improve expression efficiency, and minimize unintended effects.
 Cell Line Development: In the biopharmaceutical field, it is used to characterize the integration status of exogenous genes in engineered cell lines (such as CHO cells) to ensure stable and efficient expression of the target protein.
 Basic Scientific Research: In functional genomics research, Integration Site Analysis helps elucidate gene function and regulatory mechanisms.
Exogenous gene Integration Site Analysis services are an indispensable quality control and safety assessment tool in the field of genetic engineering. Through precise and comprehensive integration site analysis, they not only ensure the safety of gene therapy and genetically modified organisms but also provide critical data support for scientific research and industrial applications. With the continuous advancement of technology, this service will play an increasingly important role in promoting innovation and development in biotechnology.
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