一种基于CRISPR-Cas9蛋白质的改进方法,用于淘汰昆虫Sf9细胞基因
Miguel Graça1,2, Nikolaus Virgolini1,2, Ricardo Correia1,2
1iBET, Instituto de Biologia Experimental E Tecnológica, Apartado 12, Oeiras, 2780-901, Portugal.
Applied microbiology and biotechnology
|January 26, 2026
概括
这项研究为昆虫细胞引入了一种高效的CRISPR-Cas9基因编辑管道,增强生物制药生产. 在Sf9细胞中淘汰Sf-Dronc可以增强细胞灭亡抵抗力,并增加流感病毒样颗粒的产生.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 昆虫细胞对生物制药生产有价值,但缺乏先进的基因工程工具.
- 克里斯普尔-Cas9技术提供了强大的基因编辑能力,在昆虫细胞中应用有限.
研究的目的:
- 开发和实施一种高效的CRISPR-Cas9管道,用于Sf9昆虫细胞中的基因编辑.
- 通过敲除卡斯巴酶启动器Sf-Dronc来改造Sf9细胞,以增强亡抵抗力.
- 评估Sf-Dronc淘汰对细胞表型和各种生物制药产品生产的影响.
主要方法:
- 通过使用指导RNA和Cas9酶,为Sf9细胞建立了核蛋白 (RNP) 复合物输送策略.
- 该管道通过准fdl基因进行验证,达到68%的淘汰率.
- 使用下一代测序,针对Sf-Dronc进行了淘汰,并分析了由此产生的细胞系的亡抵抗性,活力和产品产量.
主要成果:
- 开发的CRISPR-Cas9管道表现出高效率,fdl基因的淘汰率为68%.
- 经过Sf-Dronc淘汰的工程Sf9细胞表现出对亡的抗性增加,并在感染baculovirus时延迟活力丧失.
- 删除Sf-Dronc导致流感病毒样颗粒 (VLPs) 的产生增加了一倍,同时对重组腺相关病毒 (rAAV) 和PfRipr5的产生产生最小的影响.
结论:
- 一个高效的CRISPR-Cas9基因编辑管道在Sf9昆虫细胞中成功实施.
- 经过工程改造的Sf9细胞系表现出增强的亡耐药性和改善特定生物制药的生产,特别是VLP.
- 这一进步有助于为生物制药行业开发优质的昆虫细胞宿主.
相关概念视频
CRISPR
57.8K
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
57.8K
CRISPR and crRNAs
19.0K
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
19.0K
CRISPR/Cas9 Genome Editing
1.8K
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
1.8K
Proteins: From Genes to Degradation
14.4K
Within a biological system, the DNA encodes the RNA, and the nucleotide sequence in the RNA further defines the amino acid sequence in the protein. This is referred to as “The Central Dogma of Molecular Biology” - a term coined by Francis Crick. Central dogma is a firm principle in biology that defines the flow of genetic information within any life form. The two fundamental steps in central dogma are - transcription and translation.
Transcription is the synthesis of RNA...
Transcription is the synthesis of RNA...
14.4K
Proteins: From Genes to Degradation
4.5K
4.5K
Osmoregulation in Insects
17.6K
Malpighian tubules are specialized structures found in the digestive systems of many arthropods, including most insects, that handle excretion and osmoregulation. The tubules are typically arranged in pairs and have a convoluted structure that increases their surface area.
17.6K


