Over-Expression Organoid Model for Target Profiling Analysis

The study of gene over-expression holds significant importance in biomedical research. By overexpressing target genes, their functions can be validated, signaling pathways and regulatory networks can be studied, and the mechanisms of gene expression regulation can be explored. Moreover, gene over-expression studies can uncover the roles of genes in cellular function and organ development, aiding in a deeper understanding of gene function and regulation. They also contribute to the comprehension of disease mechanisms and provide a foundation for new therapeutic strategies.

Previous gene over-expression studies have mainly relied on overexpressing cells and transgenic mice. However, both approaches fail to faithfully replicate the complex human tissue environment, limiting the in-depth investigation of gene function. To meet the needs of clients in biomedical research for models that allow a thorough understanding of target gene function and regulatory mechanisms, Creative Biolabs offers services for the construction and functional studies of over-expression organoid models.

Comprehensive Introduction to Over-expression Organoid Models

• Essence: Multicellular three-dimensional structures formed by genetically overexpressed stem cells or other cells under 3D culture conditions.
• Characteristics: The target gene is expressed in cells or tissues in an excessively overexpressed manner.
• Method: Achieved through gene editing techniques or transfection techniques.
• Purpose: To simulate the over-expression of the target gene in physiological or pathological conditions.
• Advantage: Highly mimics the structure and function of human organs or diseased tissues.

Research Approach Based on Over-expression Organoid Models

1. Organoid Construction: Construct pathological organoids, such as those exhibiting drug resistance, insensitivity, or high invasiveness.
2. Identification of Target Genes: Select suitable target genes based on research objectives, using transcriptomics or proteomics screening. These genes can be key genes within specific gene families or signaling pathways.
3. Further Screening of Target Genes: Validate the selected target genes in a large number of clinical samples to confirm their relevance, further narrowing down the target genes for study.
4. Expression Vector Design: Design appropriate expression vectors, including selecting suitable promoters, regulatory elements, and tags, to achieve the over-expression and visualization of the target genes.
5. Construction of Over-expression Organoids: Clone the target genes into the expression vectors and introduce the expression vectors into the target cells or organoids using appropriate techniques (e.g., restriction enzyme digestion and ligation, chemical transformation, electroporation, etc.).
6. Over-expression Validation: Validate the over-expression of the target genes in the organoids using appropriate methods such as PCR, Western blotting, and immunohistochemistry, ensuring the stability and consistency of their expression.
7. Mechanism Studies: Perform transcriptomic and proteomic analyses comparing the gene over-expression organoids with control organoids to elucidate downstream pathways of the target genes.
8. Problem-solving: Validate the effects using inhibitors of key genes in the identified optimal pathways, proposing potential therapeutic strategies.

Research Literature Based on Over-expression Organoid Models

Title:

Organoid modeling identifies that Dachshund homologue 1 (DACH1) functions as a tumor promoter in colorectal cancer by modulating Bone Morphogenetic Protein (BMP) signaling

Background:

The authors, in a previous study, identified high expression of DACH1 in colorectal cancer tissues through the analysis of clinical samples and found that DACH1 promotes the proliferation of colon cancer cells at the cellular and nuclear animal levels.

Model Used:

Gene-overexpressing colorectal cancer organoids with DACH1 over-expression.

Research Findings:

DACH1 over-expression promotes the proliferation of colon cancer stem cells. Subsequent transcriptomic analysis and pathway studies mutually confirmed that DACH1 induces the upregulation of stem cell characteristic genes by inhibiting the BMP signaling pathway, further confirming the positive regulation of colorectal cancer growth by DACH1.

Fig.1 DACH1 overexpression enhances the growth of organoids derived from normal colon crypts and colorectal adenomas.¹

Construction and Research Services of Over-expression Organoid Models at Creative Biolabs

• Professional Research Team
• High-Quality Data
• Customized Services
• Diverse Technological Platforms

Creative Biolabs is dedicated to assisting clients in achieving success in the construction and research of over-expression organoid models. If you have any need for over-expression organoid models, please contact us for more details.

Related Sections:

• Gene Editing Organoid Based Target Analysis and Validation Services
• Off-Target Screening Cell Microarray Assay