The potential of exosomes as an alternative delivery vector for the CRISPR/Cas9 components encourages Creative Biolabs to present its exosome engineering strategies.
A recently published article in the Journal of Nanobiotechnology made a comprehensive review of the current status of exosome-based CRISPR/Cas delivery systems and highlighted the potential of exosome-based delivery systems in developing novel gene delivery approaches and their application perspectives in gene editing technologies and gene therapies. Creative Biolabs, a dedicated exosome service and product provider with a global presence, is proud to present its exosome engineering service portfolio to advance the targeted delivery of CRISPR/Cas systems, especially solutions for exosome cargo loading.
Viral vectors, the leading tool for delivering CRISPR/Cas in vivo, have been found to have limitations in packaging constraints, immunogenicity, carcinogenesis, scale-up production, and the longevity of Cas expression. While the drawbacks of nonviral carriers lie in rapid clearance, biocompatibility, toxicity, immunogenicity, and therapeutic cargo release.
“Current biochemical and proteomic techniques can thoroughly characterize many proteins, nucleic acids, lipids, and other components transported by exosomes. For example, the mass spectrometry-based exosome characterization technique can easily obtain accurate information on the proteomic components of exosomes”. The exosome expert at Creative Biolabs said. “Through participating in several cell signaling pathway modulation, these exosomal components affect a variety of pathogenic activities. That’s why we believe exosomes are potential biomarkers and natural therapeutic agents for a variety of disorders. And we are excited to see the potential of exosomes in the field of gene delivery.”
Nanosized extracellular vesicles (EVs), including exosomes (30-150nm) and microvesicles (MVs; 50-1000 nm), are both described as possible alternative delivery vectors for CRISPR/Cas9 components due to high biocompatibility and stability. However, extracellular vesicles with different sizes may trigger unique uptake mechanisms. Creative Biolabs offers exosome characterization based on TRPS technology, which enables high-throughput measurement of size distribution and concentration of different extracellular vesicles to achieve higher rates of effective intracellular delivery.
Though current studies indicate exosomes are an effective tool for CRISPR/Cas9 component delivery, natural exosomes present limitations in their transportation into target sites due to exogenous and endogenous barriers. Emerging strategies can improve the loading capacity, safety, stability, targeting, and tracking of engineered exosomes for targeted delivery of the CRISPR/Cas system. Strategies for engineering exosomes include pre-loading methods, post-loading methods, and other loading methods. Creative Biolabs covers comprehensive pre- and post-loading services for exosomes to load molecules in/on exosomes:
- Introduce cargo into isolated and purified exosomes via co-incubation.
- Apply current to rearrange the phospholipid bilayer of exosomes and form small pores in the membrane, through which drugs and/or nucleotides are diffused into exosomes.
- Use ultrasonic frequencies to agitate the particles in the suspension and the probe sonicator to induce plasma membrane deformation, after which the cargo diffuses inside the exosomes.
- Incubate exosomes with the cargo and repeat freeze-thaw cycles to destroy the plasma membrane, so that the drug molecules can diffuse inside the exosomes.
- Incubate exosomes with cargos and saponin to increase permeability to ease the loading of hydrophilic molecules.
- Transfect/infect parental cells with DNA encoding therapeutically active compounds and release them via exosomes.
- Load parental cells with small molecule chemical drugs with low cytotoxicity and release them in exosomes.
The CRISPR/Cas system is a revolutionary genome editing tool, and an appropriate carrier is a precondition for ensuring the safety, efficacy, and specificity when delivering the CRISPR/Cas components to the target sites. The use of EVs in CRISPR/Cas delivery systems is still in its early stages, so researchers at Creative Biolabs are delighted to discuss and develop different design, modification, or engineering strategies and witness advanced, significant applications of exosomes in gene editing.
Creative Biolabs (https://www.creative-biolabs.com/exosome/), headquartered in New York, is a reputed contract research organization specializing mainly in exosome solutions in the U.S. and around the world. Creative Biolabs is an established player in the exosome service and product market emphasizing upstream and downstream applications of exosomes. Creative Biolabs has a complete exosome-based solution portfolio, including exosome sampling, analysis, and manufacturing.