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Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9
Published on: April 10, 2018
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CRISPR-Cas9 HDR system enhances AQP1 gene expression
Zhimin Wang1, Yaohe Wang1,2, Songling Wang3
1Sino-British Research Centre for Molecular Oncology, National Centre for International Research in Cell and Gene Therapy, School of Basic Medical Sciences, Academy of Medical Sciences, Zhengzhou University, Zhengzhou 450052, China.
Oncotarget
|January 18, 2018
Summary
Gene editing using CRISPR-Cas9 successfully increased aquaporin 1 (AQP1) expression in salivary cells. This approach shows promise for treating radiation-induced xerostomia (dry mouth).
Area of Science:
- Biotechnology
- Molecular Biology
- Genetics
Background:
- Ionizing radiation (IR) is a primary treatment for head and neck cancers.
- IR often causes irreversible salivary gland dysfunction (xerostomia), with no effective treatments.
- Aquaporin 1 (AQP1) gene therapy is a potential strategy to restore salivary function.
Purpose of the Study:
- To investigate the use of CRISPR-Cas9 gene editing to enhance aquaporin 1 (AQP1) expression.
- To assess the potential of AQP1 gene therapy for treating xerostomia.
Main Methods:
- CRISPR-Cas9 gene editing was employed in HEK293 and MDCK cell lines.
- The endogenous promoter of AQP1 was targeted for replacement with the cytomegalovirus (CMV) promoter.
- Functional assays were performed to measure transmembrane fluid flux.
Main Results:
- Successful integration of the CMV promoter led to significantly increased AQP1 gene transcription and translation.
- Overexpression of AQP1 enhanced transmembrane fluid flux, indicating improved intracellular fluid movement.
- Off-target analysis confirmed high specificity of the Cas9 cleavage for the targeted AQP1 gene.
Conclusions:
- Endogenous promoter replacement using CRISPR-Cas9 is a viable method to boost AQP1 expression.
- This gene editing strategy holds potential as a therapeutic approach for salivary gland dysfunction and xerostomia.
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