Bi-functionalized aminoguanidine-PEGylated periodic mesoporous organosilica nanoparticles: a promising nanocarrier for delivery of Cas9-sgRNA ribonucleoproteine
Abstract:Background
There is a great interest in the efficient intracellular delivery of Cas9-sgRNA ribonucleoprotein complex (RNP) and its possible applications for in vivo CRISPR-based gene editing. In this study, a nanoporous mediated gene-editing approach has been successfully performed using a bi-functionalized aminoguanidine-PEGylated periodic mesoporous organosilica (PMO) nanoparticles (RNP@AGu@PEG1500-PMO) as a potent and biocompatible nanocarrier for RNP delivery.
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“…This temporal control enables on-demand and transient editing of genomes, whereas quantitative control only targets genome editing of specific organs or tissues, reducing off-target effects and increasing immunogenicity due to long-term expression. 23,34 For instance, temperature-sensitive polymer delivery systems have been reported to achieve temporal and spatial control by physiological temperature stimulation triggering the polymer to form a gel and followed by a slow release of CRISPR/Cas9, suggesting that stimulus-triggered polymer carriers can enhance cellular uptake, lysosomal escape, intracellular precision targeting, and efficiency in delivery for CRISPR/Cas9 genomic editing systems. 35,36 In addition, it is noteworthy that stimulus-responsive polymers with a high loading capacity have recently been reported to form nanoparticles or hydrogels that induce stronger and longer-lasting immune cell responses, outperforming other non-viral carriers, with promising applications in immunotherapy of cancer and genetic diseases.…”
Section: Structural Designs Of Stimulusresponsive Polymers For Delive...mentioning
confidence: 99%
“…14,15 Non-viral delivery vectors (inorganic nanoparticles, liposomes and polymers) have significant advantages over physical methods (electroporation, microinjection and microfluidics), [16][17][18][19] viral vectors (adenoviral vectors (AV), adeno-associated viruses (AAV), retroviral and lentiviral vectors (LV)), and cationic liposomal amines for delivery of the gene editing systems of CRISPR/ Cas9. [20][21][22][23][24][25] Polymer based delivery vectors could be modified specifically to target cell-or tissue-specific responses (i.e., pH, redox, and enzymes) to enhance the organ/tissue specificity through temporally and spatially controlled CRISPR/ Cas9 genome editing, which not only has low immunogenicity and good biocompatibility, but also overcomes the packaging limitations of viral vectors, reduces the oncogenic risk, and facilitates large-scale production. [26][27][28][29] Accordingly, stimuli-responsive polymer delivery systems that are efficient and reliable as well as possess excellent targeting and superior encapsulation capabilities are currently among the major advances in genome editing systems with CRISPR/Cas9.…”
Non-viral polymeric vectors with good biocompatibility have been recent explored as delivery systems for clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) nucleases. In this review, based on current limitations...
“…This temporal control enables on-demand and transient editing of genomes, whereas quantitative control only targets genome editing of specific organs or tissues, reducing off-target effects and increasing immunogenicity due to long-term expression. 23,34 For instance, temperature-sensitive polymer delivery systems have been reported to achieve temporal and spatial control by physiological temperature stimulation triggering the polymer to form a gel and followed by a slow release of CRISPR/Cas9, suggesting that stimulus-triggered polymer carriers can enhance cellular uptake, lysosomal escape, intracellular precision targeting, and efficiency in delivery for CRISPR/Cas9 genomic editing systems. 35,36 In addition, it is noteworthy that stimulus-responsive polymers with a high loading capacity have recently been reported to form nanoparticles or hydrogels that induce stronger and longer-lasting immune cell responses, outperforming other non-viral carriers, with promising applications in immunotherapy of cancer and genetic diseases.…”
Section: Structural Designs Of Stimulusresponsive Polymers For Delive...mentioning
confidence: 99%
“…14,15 Non-viral delivery vectors (inorganic nanoparticles, liposomes and polymers) have significant advantages over physical methods (electroporation, microinjection and microfluidics), [16][17][18][19] viral vectors (adenoviral vectors (AV), adeno-associated viruses (AAV), retroviral and lentiviral vectors (LV)), and cationic liposomal amines for delivery of the gene editing systems of CRISPR/ Cas9. [20][21][22][23][24][25] Polymer based delivery vectors could be modified specifically to target cell-or tissue-specific responses (i.e., pH, redox, and enzymes) to enhance the organ/tissue specificity through temporally and spatially controlled CRISPR/ Cas9 genome editing, which not only has low immunogenicity and good biocompatibility, but also overcomes the packaging limitations of viral vectors, reduces the oncogenic risk, and facilitates large-scale production. [26][27][28][29] Accordingly, stimuli-responsive polymer delivery systems that are efficient and reliable as well as possess excellent targeting and superior encapsulation capabilities are currently among the major advances in genome editing systems with CRISPR/Cas9.…”
Non-viral polymeric vectors with good biocompatibility have been recent explored as delivery systems for clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) nucleases. In this review, based on current limitations...
“…More recently, editing genetic information in mammalian cells using CRISPR/Cas9 has shown great potential in developing a new generation of protein therapeutics for the treatment of genetic diseases. 259 Similarly, Chang et al screened several major nanoparticle formulations based on a combinatorial library of cationic lipid for intracellular protein delivery. 260 In this Account, the researcher optimized the chemical structure of lipids to control lipid degradation and release of intracellular proteins for delivery of CRISPR/Cas9 genome editing proteins.…”
Intracellular cargo delivery, the introduction of small molecules, proteins, and nucleic acids into a specific targeted site in a biological system, is an important strategy for deciphering cell function, directing...
“…For example, Waggoner et al improved loading of a therapeutic protein by >50-fold 13% w/w (∼50% v/v) in PSiNPs vs <1% w/w (<1% v/v) in PLGA] . Although no one has yet demonstrated Cas9 RNP delivery with PSiNPs, silica nanoparticles have been demonstrated for gene editing. − While the route of synthesis, geometry, and chemical structure are very different between PSi and silica, both can achieve high drug loading (∼10% w/w).…”
The complexity of CRISPR machinery is a challenge to
its application
for nonviral in vivo therapeutic gene editing. Here,
we demonstrate that proteins, regardless of size or charge, efficiently
load into porous silicon nanoparticles (PSiNPs). Optimizing the loading
strategy yields formulations that are ultrahigh loading>40%
cargo by volumeand highly active. Further tuning of a polymeric
coating on the loaded PSiNPs yields nanocomposites that achieve colloidal
stability under cryopreservation, endosome escape, and gene editing
efficiencies twice that of the commercial standard Lipofectamine CRISPRMAX.
In a mouse model of arthritis, PSiNPs edit cells in both the cartilage
and synovium of knee joints, and achieve 60% reduction in expression
of the therapeutically relevant MMP13 gene. Administered intramuscularly,
they are active over a broad dose range, with the highest tested dose
yielding nearly 100% muscle fiber editing at the injection site. The
nanocomposite PSiNPs are also amenable to systemic delivery. Administered
intravenously in a model that mimics muscular dystrophy, they edit
sites of inflamed muscle. Collectively, the results demonstrate that
the PSiNP nanocomposites are a versatile system that can achieve high
loading of diverse cargoes and can be applied for gene editing in
both local and systemic delivery applications.
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