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关注球盟会生物基因,可及时获取更多优惠内容!
CRISPR那些事儿
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FAQ
How does EDITGENE’s Bingo™ Prime Editing 7 (PE7) platform ensure point mutation success rates?
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.
How is Prime Editing 7 (PE7) different from traditional CRISPR/Cas9 technology?
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.
Are all types of genes suitable for KO cell lines?
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.
Why choose EDITGENE?
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.
What is the difference between KO cell lines and gene knockout animal models?
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.
Why do researchers use KO cell lines?
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.
What is a KO cell line?
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.
Are KO cell lines applicable to all cell types?
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.
What is gene overexpression?
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.
Why conduct gene overexpression?
Gene overexpression aids in studying the function of specific genes, revealing their role within the organism. It is also commonly used in drug screening, vaccine development, and protein production. For example, by overexpressing a therapeutic protein, researchers can evaluate its efficacy in disease models.
为什么选择球盟会生物基因KO细胞系?
球盟会生物基因KO细胞库有4500+株高质量KO细胞系现货可供选择,可大幅度节省您的科研时间。另外球盟会生物基因可提供高效、高阳性率的基因敲除定制服务,全球知名院校博士团队一对一服务。
KO细胞系是否适合所有类型的基因?
虽然KO细胞系技术强大,但并非所有基因都适合敲除。某些基因的敲除可能导致细胞死亡或严重的生理功能障碍,特别是那些对细胞存活至关重要的基因。在这种情况下,可以选择条件性敲除或基因敲降(如RNAi)来研究这些基因的功能。
为什么研究人员要使用KO细胞系?
研究人员使用KO细胞系来解析基因功能,因为敲除基因能够揭示该基因在细胞生长、分化、代谢、信号传导等过程中的具体作用。顺利获得比较敲除基因的细胞与正常细胞的差异,科研家可以深入分析基因在疾病中的作用,进而开发新的治疗方法。例如,KO细胞系在癌症、遗传病和神经退行性疾病的研究中具有重要应用。
一般CRISPR相关试剂以及Cas蛋白可以存放多久?
CRISPR检测相关试剂:
①RPA恒温扩增试剂盒于-20℃保存,可长期存放。
②靶标质粒可长期存放-20℃ ③ Cas蛋白反复冻融容易影响活性,建议分装多管,存放-80℃,根据实验需要取出使用。另外,短期内使用的,可放-20℃保存。
③crRNA容易降解,建议短时间内用不到的于-80℃保存。
④探针是DNA双链,相对来说没有那么容易降解,可放-20℃保存。
①RPA恒温扩增试剂盒于-20℃保存,可长期存放。
②靶标质粒可长期存放-20℃ ③ Cas蛋白反复冻融容易影响活性,建议分装多管,存放-80℃,根据实验需要取出使用。另外,短期内使用的,可放-20℃保存。
③crRNA容易降解,建议短时间内用不到的于-80℃保存。
④探针是DNA双链,相对来说没有那么容易降解,可放-20℃保存。
如何选择全基因组或亚基因组CRISPR文库?
CRISPR文库类型可分为全基因组文库和亚基因组文库。如果目标是进行全基因组范围内的筛选,那么全基因组文库是最佳选择。这类文库通常包含针对整个基因组的sgRNA。如果研究目标有偏向性,如只关注特定基因家族或特定信号通路,那么可以选择亚基因组文库,以减少不必要的筛选工作量和成本。