Lentivirus is a gene delivery tool that introduces exogenous genes into cells. By using tool cells such as 293T, lentiviral vectors carrying target DNA fragments are packaged into lentiviral particles with cell-infectious activity. This packaging process includes constructing lentiviral vectors, preparing packaging plasmids, culturing tool cells, transfecting plasmids, collecting viral particles, purifying and concentrating viral particles, and titration.

Lentiviruses have high transduction efficiency and the ability to maintain stable gene expression over the long term, making them particularly suitable for cell types that are difficult to transfect. Furthermore, lentiviruses can integrate exogenous genes into the host genome, ensuring prolonged gene expression.

4.Poor cell condition after lentiviral infection may be caused by various factors. Here are some possible reasons and corresponding solutions: 1.High viral titer: High titers of lentivirus may cause cytotoxicity, preventing normal cell growth. Solution: Lower the viral titer and conduct a series of dilution experiments to find a titer that effectively transduces without adversely affecting cell growth. 2.Poor cell condition: The health status of cells before infection can affect growth after infection. Solution: Ensure cells are in optimal condition for infection, for example, by changing to fresh culture medium 24 hours before infection and ensuring appropriate cell density. 3.Toxicity of gene expression mediated by the virus: The gene carried by the lentiviral vector may be toxic to the cells, affecting their growth. Solution: If possible, use a control vector to determine if the problem is related to gene expression, and select appropriate vectors or genes for research. 4.Excessive antibiotic selection pressure: If antibiotics are used to select transfected cells, excessive concentrations of antibiotics may inhibit cell growth. Solution: Optimize the antibiotic concentration and use gradient experiments to determine the optimal concentration.

EDITGENE’s newly upgraded seventh-generation Bingo™ Prime Editing (PE7) platform optimizes editing protein and RNA editing activity. Compared to the fifth-generation PE technology, point mutation success rates and gene editing efficiency have significantly improved, with one-on-one support from PhDs from globally renowned institutions.

Prime Editing is a novel gene editing technology that enables precise gene editing without introducing double-strand DNA breaks. It has two core components: pegRNA and the PEmax gene-editing enzyme (Cas9n-RT). PegRNA not only targets the desired sequence but also contains the base modification information. In the editing system, pegRNA guides PEmax to the designated edit site, nicks the DNA single strand, and reverse transcribes the sequence within the pegRNA to modify, inserting it into the target genome location, thereby achieving precise single-base substitutions or small insertions and deletions

EDITGENE’s Bingo™ Prime Editing 7 (PE7) platform is built upon over ten years of gene editing experience, with optimization and advancements derived from thousands of gene editing CRO projects, achieving significantly higher success rates than traditional site-specific mutation systems. The Bingo™ Prime platform utilizes highly efficient reverse transcriptase and precise guide RNA design, ensuring each point mutation reaches the desired outcome.

Traditional CRISPR/Cas9 technology achieves gene editing by introducing double-strand breaks at the target DNA site and then using the cell’s homologous recombination repair mechanism. This approach carries multiple risks, such as lower editing efficiency, reduced homozygous mutation rates, and random insertions or deletions. Prime Editing, however, does not require double-strand breaks. With its Cas9n-RT editing enzyme system and pegRNA, Prime Editing achieves more accurate and safer gene editing with reduced off-target effects.

Not all genes are suitable for knockout. Some gene knockouts may result in cell death or severe dysfunction, particularly for essential genes. In such cases, conditional knockouts or gene knockdowns (e.g., RNAi) may be used instead.

EDITGENE provides access to a comprehensive library of over 4,500 high-quality knockout (KO) cell lines, enabling researchers to save valuable time. Our custom gene knockout services are highly efficient, boasting a high positive rate while minimizing off-target effects. Clients also benefit from personalized, one-on-one support from a team of PhD experts from globally renowned institutions, ensuring top-tier service and results.

KO cell lines are used for in vitro experiments, suitable for high-throughput screening and cellular studies, while gene knockout animal models are used for in vivo experiments to study gene functions within an entire organism and its interaction with the environment.

Researchers use KO cell lines to investigate gene functions by observing the effects of gene deletion on cellular behavior. This helps in understanding the role of genes in various processes like cell growth, metabolism, and signal transduction. KO cell lines are vital for studying diseases like cancer, genetic disorders, and neurodegenerative diseases.

KO (Knockout) cell line is a cell line where a specific gene has been completely removed or rendered non-functional through gene editing technologies such as CRISPR-Cas9. These cell lines are critical for understanding gene functions and disease mechanisms.

KO cell lines can be applied to various cell types, including cancer cells, stem cells, and primary cells, but different cell types may have varying sensitivities to gene editing, and may vary among different cell types. In certain cell types, achieving gene knockout may require optimization of transfection conditions and selection of appropriate gene-editing tools.

Gene overexpression refers to using various techniques to significantly increase the expression level of a specific gene in cells or organisms. This is often achieved by introducing additional gene copies or using strong promoters to drive gene expression.