Related Experiment Video
Updated: Jun 7, 2025
09:51
Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
Published on: May 25, 2018
33.8K
Rationally designed Campylobacter jejuni Cas9 enables efficient gene activation and base editing
Yuxi Chen1, Rui Kang1, Yuanling Jiang1,2
1MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
Molecular Therapy. Nucleic Acids
|November 19, 2024
Summary
Researchers engineered the compact CRISPR-Cas9 system (CjCas9) for improved genome editing. Enhanced CjCas9, delivered via adeno-associated virus (AAV), shows high efficiency and safety for therapeutic applications.
Area of Science:
- Molecular Biology
- Gene Editing Technologies
- Biotechnology
Background:
- CRISPR-Cas systems are crucial for genome engineering.
- Adeno-associated virus (AAV) is a common delivery vector.
- The compact CjCas9 offers advantages for AAV delivery but requires efficiency improvements.
Purpose of the Study:
- To enhance the editing efficiency of the CjCas9 system.
- To develop novel CRISPR-based tools for gene modulation and base editing.
- To evaluate the in vivo efficacy and safety of engineered CjCas9 systems.
Main Methods:
- Structure-guided protein engineering of CjCas9.
- Development of a miniature transcriptional activator (LDE-CjCas9-VPR).
- Engineering of CjCas9-derived base editors (LDE-CjABE, LDE-CjCBE).
- In vitro editing in human and mouse cells.
- In vivo AAV-mediated delivery and analysis in mouse retina.
Main Results:
- Engineered CjCas9 variants demonstrated improved editing efficiency.
- LDE-CjABE successfully induced genome editing in human and mouse cells.
- AAV delivery of LDE-CjABE enhanced on-target editing efficiency in vivo.
- No off-target editing was detected in the mouse retina.
Conclusions:
- The engineered LDE-CjCas9 system significantly improves editing efficiency.
- Compact and efficient CjCas9 tools expand therapeutic genome editing applications.
- AAV-delivered LDE-CjCas9 variants show promise for safe and effective gene therapy.
Related Concept Videos
01:59CRISPR
49.5K
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...
49.5K
02:53CRISPR and crRNAs
16.8K
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...
16.8K
02:31Homologous Recombination
50.2K
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...
50.2K
02:53Conservative Site-specific Recombination and Phase Variation
5.9K
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...
5.9K