高通量 (PbHiT):一种CRISPR-Cas9系统,用于大规模研究基因
Thorey K Jonsdottir1,2, Martina S Paoletta1,2,3, Johan Henriksson2,4,5,6
1The Laboratory for Molecular Infection Medicine Sweden, Umeå University, Umeå, Sweden.
Bio-protocol
|January 29, 2026
概括
菌高通量 (PbHiT) 系统提供了一个可扩展的CRISPR-Cas9方法,用于有效编辑疟疾寄生虫的基因组. 这种工具可以快速生成和识别淘汰和标记的寄生虫线,用于功能研究.
科学领域:
- 分子生物学分子生物学
- 寄生虫学的寄生虫学
- 遗传学 是一个遗传学.
背景情况:
- 基因改造对于了解菌寄生虫生物学至关重要,但由于基因可处理性低,依赖同源重组,因此面临挑战.
- 像PlasmoGEM这样的现有方法在覆盖范围和灵活性上有局限性,用于全基因组淘汰.
研究的目的:
- 开发一种可扩展的CRISPR-Cas9协议,用于在Plasmodium berghei中有效编辑基因组.
- 创建一个高吞吐量系统,用于产生淘汰和标记的寄生虫线.
- 为动物疟疾寄生虫的功能研究和遗传查提供可复制的工具.
主要方法:
- 菌高通量 (PbHiT) 系统使用单个克隆步骤将指导RNA (gRNA) 与短同质臂 (100 bp) 连接起来.
- 聚合载体转染被gRNA条形码启用,允许通过下游gRNA测序识别突变物.
- 该协议支持对基因组编辑的淘汰和标记策略.
主要成果:
- PbHiT系统证明了P. berghei. 的高效和可复制的基因组编辑.
- 它可靠地回顾了已知的突变生长表型.
- 该方法支持高吞吐量产生淘汰和标记的寄生虫线.
结论:
- PbHiT系统为P. berghei基因组编辑提供了一个可扩展和高效的CRISPR-Cas9平台.
- 该协议促进了有针对性的功能研究和高通量遗传选.
- 一个P. berghei基因构造设计的在线资源可用于支持该协议.
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
19.1K
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...
19.1K
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
pH Scale
79.6K
Hydronium and hydroxide ions are present both in pure water and in all aqueous solutions, and their concentrations are inversely proportional as determined by the ion product of water (Kw). The concentrations of these ions in a solution are often critical determinants of the solution’s properties and the chemical behaviors of its other solutes. Two different solutions can differ in their hydronium or hydroxide ion concentrations by a million, billion, or even trillion times. A common means of...
79.6K
Scaling
593
In designing and analyzing filters, resonant circuits, or circuit analysis at large, working with standard element values like 1 ohm, 1 henry, or 1 farad can be convenient before scaling these values to more realistic figures. This approach is widely utilized by not employing realistic element values in numerous examples and problems; it simplifies mastering circuit analysis through convenient component values. The complexity of calculations is thereby reduced, with the understanding that...
593
Gene Families
9.9K
Gene families consist of groups of genes proposed to have originated from a common ancestor. Typically these arise through events in which a gene or genes are mistakenly duplicated during cell division. Unlike their parent genes (which are subject to selection pressure to maintain function), these gene copies do not need to preserve their sequences and may evolve at a relatively faster rate.
Occasionally these regions can be adapted to take on new roles within the organism, becoming novel genes...
Occasionally these regions can be adapted to take on new roles within the organism, becoming novel genes...
9.9K


