相关实验视频

Updated: Jul 4, 2025

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Published on: October 18, 2022

通过使用混合dCas12a变体和发针间隔器crRNAs进行精确和多重基因调节的工程CRISPRa系统

Yuqing Ke^1,2, Shuang Zhang^1,2, Yingying Gao^1,2

¹Nantong First People's Hospital and Nantong Hospital of Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Nantong 226006, P. R. China.

Analytical chemistry

|February 2, 2024

概括

研究人员改进了CRISPR-Cas12a基因编辑,开发了一种混合Cas12a变体和发针间隔crRNAs (h-CAP). 这种增强的系统为人类细胞中的基因表达调节提供了更大的特异性和效率.

科学领域:

分子生物学分子生物学
基因编辑技术的技术
生物技术是生物技术.

背景情况:

CRISPR-Cas12a是一种基因编辑工具,在基因编辑和表达调节方面具有潜在的应用.
目前Cas12a的局限性包括非目标效应和效率降低,阻碍其广泛采用.
开发提高Cas12a特异性和效率的策略对于推进基因编辑应用至关重要.

研究的目的:

为了提高CRISPR-Cas12a系统的特异性和效率.
开发一种使用混合工程 Cas12a 变体和发针间隔 crRNAs (h-CAP) 的新方法.
证明h-CAP策略对于精确基因表达调节的能力.

主要方法:

使用了混合工程 Cas12a 变种.
用发针间隔crRNAs (h-CAP) 来进行向基因操纵.
在人类细胞中测试了该系统的性能,以调节基因表达.

主要成果:

使用h-CAP策略,证明了CRISPR-Cas12a系统的增强特异性.
在基因编辑和表达调节方面实现了提高效率.
在人类细胞中成功实施了单基特异性和多重基因表达调节.

结论:

混合工程 Cas12a 变种与 h-CAP 结合,显著提高了 CRISPR-Cas12a 系统的特异性和效率.
这种新的方法可以实现精确的单基因特异性和多基因基因表达调节.
在基因编辑和合成生物学中,h-CAP战略代表了CRISPR-Cas12a应用的有希望的进步.

更多相关视频

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

Published on: September 2, 2021

Efficient Generation of hiPSC Neural Lineage Specific Knockin Reporters Using the CRISPR/Cas9 and Cas9 Double Nickase System

Efficient Generation of hiPSC Neural Lineage Specific Knockin Reporters Using the CRISPR/Cas9 and Cas9 Double Nickase System

Published on: May 28, 2015

相关概念视频

CRISPR

CRISPR

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...

CRISPR and crRNAs

CRISPR and crRNAs

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...

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

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...

Homologous Recombination

Homologous Recombination

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...