酵母基因编辑

服务介绍

CRISPR/Cas9/gRNA（CRISPR-Cas RNA-gudide nuclease）基因敲除技术是继TALEN基因敲除技术之后又一大突破。与TALEN和ZNF技术相比，CRISPR/Cas9系统的载体构建和使用更加方便。基因敲除的特异性由gRNA而不是蛋白来确定，因此避免了复杂的质粒组装构建过程，甚至使同时敲除多个基因成为可能。不仅如此，实验表明CRISPR/Cas9系统的基因敲除效率与TALEN、ZNF的效率相当或者更好。目前Cas9的特异性及其它一些特性尚有待进一步研究，但由于其操作极其简便，该技术的应用将大大简化育种，遗传变异修复，及各种基因研究的工作难度。

服务优势

▶多经验丰富：在酵母基因编辑领域有丰富的经验积累，经验丰富的团队可以帮助您快速完成酵母突变体构建；

▶两种实验方案：无痕/重组；

▶任意位点：使用我们先进的Cas9技术，可在任何位点引入突变；

▶较长DNA引入：经过技术优化提升，可在长达10 kb的DNA基因敲入；

▶一站式解决方案：提供全面的上下游服务，包括酵母基因敲除及酵母回补实验验证；

▶质量保证：确保所有基因编辑位点都经过测序鉴定，以确保产品质量。

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文案展示

An Introduced RNA-Only Approach for Plasmid Curing via the CRISPR-Cpf1 System in Saccharomyces cerevisiae

期刊：Biomolecules

evelopment of a plasmid curing system using CRISPR-Cpf1 system in S. cerevisiae. (a) Schematic representation of the plasmid pBCscADE2, crRNAs targeting for the replication origin and the homologous DNA template of CEN/ARS sequence. (b) Plasmid-cured colonies can grow on YPD medium without G418, but not on medium containing G418. The arrows indicate the colonies which failed plasmid curing. (c) Colony PCR by primers specific for the plasmid. (d) The

plasmid-curing efficiency for crCEN1 and crCEN2. Strains were transformed crCEN1 or crCEN2 with CEN HA donor DNA and plated on YPD medium with or without G418. Data were derived from three separate experiments and are presented as the means ± SD. The symbol * indicated statistically significant differences (p < 0.05 using Student’s t-test).

Figure 1. Development of a plasmid curing system using CRISPR-Cpf1 system in S. cerevisiae.(a) Schematic representation of the plasmid pBCscADE2, crRNAs targeting for the replication origin and the homologous DNA template of CEN/ARS sequence. (b) Plasmid-cured colonies can grow on YPD medium without G418, but not on medium containing G418. The arrows indicate the colonies which failed plasmid curing. (c) Colony PCR by primers specific for the plasmid. (d) The plasmidcuring efficiency for crCEN1 and crCEN2. Strains were transformed crCEN1 or crCEN2 with CEN HA donor DNA and plated on YPD medium with or without G418. Data were derived from three separate experiments and are presented as the means ± SD. The symbol * indicated statistically significant differences (p < 0.05 using Student’s t-test).

适用领域

▶蛋白质功能机制研究：是解析蛋白质结构与功能关系的核心工具。通过对将待研究基因转入相关酵母突变体，观察蛋白活性、结合能力、亚细胞定位等变化，明确蛋白的功能作用，揭示蛋白质发挥生理功能的分子机制。

▶工业生物制造：酵母被广泛用于可持续生产高价值的化学品和材料。通过对酵母进行基因编辑和代谢工程改造，可以使其生产多种产品。

▶工业菌株性能优化：基因编辑可以系统性提升酵母在工业生产中的综合性能，如提升耐受性，以增强菌株对高温、毒素等恶劣条件的抵抗能力。再次，优化代谢通路，获得生产能力更强的工业菌株。

▶生物医药与重组蛋白：以毕赤酵母为代表的表达系统是生产复杂重组蛋白的重要平台。基因编辑技术在其中扮演关键角色。构建高效菌株：通过敲除特定基因（如KU70、pep4），提升外源蛋白的稳定性和产量。优化表达系统：利用CRISPR等技术构建诱导型表达系统，实现更安全、高效的蛋白生产。