Related Experiment Video
Updated: Sep 9, 2025
08:20
A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
Published on: September 2, 2021
4.2K
Topology-Engineered Guide RNAs for Programmable Control of CRISPR/Cas Activity
Liang Cheng1,2
1Beijing National Laboratory for Molecular Sciences (BNLMS), CAS Key Laboratory of Molecular Recognition and Function, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences, Beijing, 100190, China.
Angewandte Chemie (International Ed. in English)
|September 4, 2025
Summary
Topology-engineered guide RNAs (TE-gRNAs) offer precise control over CRISPR gene editing. These advanced RNA structures enable conditional and reversible editing, overcoming limitations of traditional methods for enhanced applications.
Area of Science:
- Molecular Biology
- Biotechnology
- Chemical Engineering
Background:
- CRISPR/Cas systems offer powerful genome editing capabilities.
- Achieving precise temporal and conditional control of CRISPR remains a significant challenge.
- Traditional linear guide RNAs (gRNAs) present limitations in control, efficiency, and reversibility.
Purpose of the Study:
- To introduce and review topology-engineered guide RNAs (TE-gRNAs) as an advanced solution for controlled CRISPR/Cas genome editing.
- To highlight the structural diversity and functional advantages of TE-gRNAs.
- To discuss the potential of TE-gRNAs in various biological and therapeutic applications.
Main Methods:
- Engineering defined RNA topologies (polymeric, circular, dendrimer-like) for gRNAs.
- Incorporating stimuli-responsive linkers and groups for external trigger control (e.g., light, chemical signals).
- Evaluating TE-gRNAs for improved synthesis, stability, reduced off-target effects, and precise spatiotemporal control.
Main Results:
- TE-gRNAs enable precise spatial and temporal control over CRISPR/Cas activity.
- Defined topologies allow for reversible and programmable activation/deactivation of editing.
- Engineered structures enhance synthesis feasibility, stability, and reduce off-target effects.
Conclusions:
- TE-gRNAs represent a significant advancement in achieving dynamic and conditional genome editing.
- Their unique structural properties offer unprecedented control over CRISPR systems.
- TE-gRNAs hold broad potential for synthetic biology, functional genomics, and therapeutic interventions.
Related Concept Videos
01:59CRISPR
52.8K
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...
52.8K
01:28CRISPR/Cas9 Genome Editing
208
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...
208
02:53CRISPR and crRNAs
17.3K
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...
17.3K
02:53Conservative 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
01:23RNA Interference
26.4K
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
26.4K