Cell lines are indispensable tools in CRISPR-Cas9 technology, providing a versatile platform for gene editing, disease modeling, and therapeutic development. From immortalized cell lines to primary cells, the choice of cell line significantly impacts the efficiency, accuracy, and relevance of CRISPR-based experiments.
Key Applications of Cell Lines in CRISPR:
- Disease Modeling: CRISPR-edited cell lines enable the creation of accurate and reproducible disease models, facilitating the study of disease mechanisms and the testing of potential therapies. By introducing specific disease-causing mutations into cell lines, researchers can mimic disease phenotypes and investigate the effects of genetic alterations on cellular function.
- Therapy Production: Genetically modified cell lines are used to produce important therapies such as monoclonal antibodies and CAR-T cells. CRISPR technology enhances cell line stability, increases target protein production, and allows for the creation of universal donor cells, which are modified to reduce the chances of rejection by the host.
- Functional Genomics: CRISPR-Cas9 technology facilitates the study of gene functions and disease mechanisms. It allows researchers to create targeted gene knockouts, generate disease models, and explore potential therapeutic strategies.
Cutting-Edge Approaches:
- Live Imaging: Modifications to the gRNA sequence allow for mRNA recognition and tracking. These new applications improve existing methodologies for live imaging within cells, such as fluorescence in situ hybridization (FISH).
- CAR-T Cell Therapies: CRISPR-Cas9 technology has enhanced CAR-T therapy by enabling precise genetic edits that improve T cell functionality, persistence, and specificity. For example, CRISPR-mediated knockout of immune checkpoint molecules, such as programmed cell death protein 1 (PD-1), demonstrated promising antitumor activity of CAR-T cells by inhibiting their exhaustion and enhancing persistence in the hostile tumor microenvironment (TME).