在Saccharomyces cerevisiae中通过工程V(D) J类重组产生组合多样性
Andrew P Cazier1, Jaewoo Son1, Sreenivas Yellayi1
1School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA, USA.
Nature communications
|July 1, 2025
概括
研究人员设计了酵母来执行V(D) J重组,这是对抗体多样性的关键过程. 这一突破使得在抗体开发和使用非脊椎动物细胞的基因工程方面有了新的应用.
科学领域:
- 分子生物学分子生物学
- 免疫学 免疫学 免疫学
- 合成生物学 合成生物学
背景情况:
- V(D) J重组对于脊椎动物的抗体多样性至关重要.
- 这一过程涉及重组激活基因1和2 (RAG1和RAG2).
- 酵母,像Saccharomyces cerevisiae一样,缺乏V(D) J重组,但为抗体显示提供了优势.
研究的目的:
- 为了使酵母经历V(D) J重组的工程设计.
- 为了提高酵母在同质辅助重组的效率.
- 建立一个研究RAG1突变和产生遗传多样性的平台.
主要方法:
- 在Saccharomyces cerevisiae中将RAG1和RAG2纳入.
- 使用分裂抗生素耐药性试验来测量重组.
- 采用各种策略来优化重组率.
主要成果:
- 在工程酵母中证明了成功的同质辅助编码关节形成.
- 实现了重组率的7000倍增加,在四天后达到近1%.
- 开发了一个平台来评估RAG1突变并产生各种蛋白质和抗体碎片.
结论:
- 工程酵母可以执行V(D) J重组,扩大其实用范围超出脊椎动物.
- 开发的平台有助于研究重组机制和致病突变.
- 这项工作代表了通过在非脊椎动物细胞中随机重组的第一代遗传和表型多样性.
相关概念视频
Carrier Generation and Recombination
819
Carrier generation is the process by which electron-hole pairs (EHPs) are created within the semiconductor. In direct-bandgap semiconductors, such as gallium arsenide (GaAs), this occurs efficiently when energy absorption prompts valence electrons to leap into the conduction band, leaving behind holes.
This process is given by the generation rate G and is efficient due to the conservation of momentum between the valence band maximum and conduction band minimum.
Indirect generation involves an...
This process is given by the generation rate G and is efficient due to the conservation of momentum between the valence band maximum and conduction band minimum.
Indirect generation involves an...
819
Gene Conversion
10.0K
Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
10.0K
Combinatorial Gene Control
8.4K
Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
8.4K
Homologous Recombination
52.6K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
52.6K
Conservative Site-specific Recombination and Phase Variation
6.1K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
6.1K
Overview of Transposition and Recombination
16.1K
Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
16.1K


