通过一个增强的,可扩展的突变基因编辑器平台,加速发现耐药变体
Kristel M Dorighi1, Anqi Zhu2, Jean-Philippe Fortin3
1Department of Molecular Biology, Genentech, Inc., South San Francisco, CA 94080, USA.
Cell reports
|June 5, 2024
概括
这项研究引入了一种新的基准编辑系统,用于快速选癌症药物耐药性突变. 该技术识别了赋予针对性疗法耐药性的基因变异,加速了个性化医学研究.
科学领域:
- 基因组学就是基因组学.
- 分子生物学分子生物学
- 癌症研究 癌症研究
背景情况:
- 个性化癌症疗法依赖于瘤遗传特征,但面临着通过突变获得药物耐药性的挑战.
- 识别耐药性转移突变往往是昂贵的,资源密集的和耗时的.
研究的目的:
- 开发和优化一种新的基准编辑系统,以高效,大规模地对导致药物耐药性的基因变异进行现场查.
- 为了确定EGFR和BRAF中赋予EGFR抑制剂耐药性的特定突变.
主要方法:
- 条件激活诱导的cytidine deaminase (AID) -死Cas9 (dCas9) 基编辑系统的调整和优化.
- 使用自定义的单向导RNA (sgRNA) 库进行向基因变异查.
- 对表皮生长因子受体 (EGFR) 和v-raf小鼠肉瘤病毒性瘤基因同类B1 (BRAF) 基因的鉴定变异的分析.
主要成果:
- 与标准的BE4基础编辑器相比,优化的AID-dCas9系统展示了更大的编辑异质性和扩展的编辑足迹.
- 成功识别了EGFR和BRAF中的单个和复合变异,这些变异对EGFR抑制剂产生抗性.
- 建立了一个可扩展的平台,用于发现药物修饰变体和理解蛋白质结构功能关系.
结论:
- 开发的基础编辑系统提供了一个简单,可扩展和高效的平台,用于发现癌症抗药机制.
- 这种方法有助于识别新的治疗点和克服癌症治疗中耐药性的策略.
- 为药物耐药性的遗传基础提供了宝贵的见解,推进了个性化瘤学.
相关概念视频
00:57Antibiotic Selection
Overview
01:59CRISPR
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 Short...
02:46Genome Copying Errors
DNA replication is a well-evolved process that copies millions of base pairs with high fidelity during each cell division. Occasionally a wrong base or a long stretch of wrong bases may get added to the daughter strands. If the errors are left unchecked, cells might accumulate several mutations that might endanger their survival. Therefore, the copying errors are checked and repaired at three levels.
01:29Pharmacogenomics: Identification of New Drug Targets
Advances in genomics have profoundly influenced drug discovery by increasing both the speed and accuracy of pharmaceutical development. Pharmacogenomics, which examines how genetic variation influences drug response, facilitates the identification of novel therapeutic targets and enables patient stratification for personalized treatment. These strategies contribute to improved drug efficacy, minimized adverse effects, and more efficient clinical trial design.Mapping genetic differences...
01:16Transduction
Among the three main modes of HGT—transformation, conjugation, and transduction—transduction is unique in that it is mediated by bacteriophages, or bacterial viruses.Transduction occurs in two ways. Generalized transduction occurs during the lytic cycle of a bacteriophage infection. In this process, bacteriophages infect bacterial cells, replicate within them, and ultimately cause cell lysis, releasing newly assembled virions. Occasionally, random fragments of the bacterial genome are...
01:28CRISPR/Cas9 Genome Editing
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...