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Once the gRNA library is designed, the gRNA sequences are synthesized and cloned into a vector, typically a lentiviral or retroviral vector. These vectors allow for efficient delivery of the gRNA into the target cells. The vector also contains a gene encoding the Cas9 enzyme, as well as a selection marker (e.g., antibiotic resistance) to allow for selection of cells that have been successfully transduced with the virus. After cloning, the vector is packaged into viral particles in specialized packaging cells. The resulting virus stock contains a diverse pool of viral particles, each carrying a different gRNA. The virus is then purified and titered to determine the concentration of infectious particles.
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* **To Maintain a Formal Tone:** In certain formal contexts, like academic writing or legal documents, passive voice can be preferred because it sounds more objective and detached. For example, “The proposal was reviewed by the committee.” This is a key difference to understand. The tone changes dramatically.
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* **Une œuvre d'art du cœur (A work of art from the heart):** This means that the artwork was created with passion and genuine emotion. It suggests that the artist poured their heart and soul into the piece, and that the work reflects their innermost feelings. The term implies that it is more than just a piece of art; it is a profound expression of self.
