Lipid Nanoparticle-Mediated Delivery of CRISPR-Cas9 for Therapeutic Gene Correction DOI Open Access
Johanna Walther

Опубликована: Ноя. 28, 2023

The thesis “Lipid Nanoparticle-Mediated Delivery of CRISPR-Cas9 for Therapeutic Gene Correction” delves into the potential as a gene therapy tool treating hereditary disorders. Originally bacterial defense mechanism, has been modified precision editing tool. It functions like molecular scissors, creating specific DNA breaks that trigger repair processes, which, when coupled with DNA, can correct mutations causing diseases. However, challenge lies in safely and efficiently delivering to affected organs patients. explores use lipid nanoparticles, small, virus-sized particles composed naturally occurring body fats, viable delivery system CRISPR-Cas9. work this demonstrates designed nanoparticles effectively transport both cultured cells later liver mice well. Moreover, results show delivered generates precise genome edits. Furthermore, it addresses immune responses against CRISPR-Cas9, given origin, introduces strategy mitigate such reactions. In summary, presents promising approach deliver diseased cells, enabling correction pathogenic mutations, marking significant advancement field

Язык: Английский

Adenine base editing of CFTR using receptor targeted nanoparticles restores function to G542X cystic fibrosis airway epithelial cells DOI Creative Commons

I Röse,

Miriam Greenwood, Matthew Biggart

и другие.

Cellular and Molecular Life Sciences, Год журнала: 2025, Номер 82(1)

Опубликована: Апрель 7, 2025

Язык: Английский

Процитировано

0
Lipid nanoparticle-encapsulated DOCK11-siRNA efficiently reduces hepatitis B virus cccDNA level in infected mice DOI Creative Commons
Hikari Okada, Takeharu Sakamoto, Kouki Nio

и другие.

Molecular Therapy — Methods & Clinical Development, Год журнала: 2024, Номер 32(3), С. 101289 - 101289

Опубликована: Июнь 24, 2024

The hepatitis B virus (HBV) infects many people worldwide. As HBV infection frequently leads to liver fibrosis and carcinogenesis, developing anti-HBV therapeutic drugs is urgent. Therapeutic for preventing covalently closed circular DNA (cccDNA) production, which can eliminate infection, are unavailable. host factor dedicator of cytokinesis 11 (DOCK11) involved in the synthesis maintenance cccDNA

Язык: Английский

Процитировано

4
Glucocorticoid pre-administration improves LNP-mRNA mediated protein replacement and genome editing therapies DOI
Li Li, Mei Luo, Lifang Zhou

и другие.

International Journal of Pharmaceutics, Год журнала: 2025, Номер unknown, С. 125282 - 125282

Опубликована: Янв. 1, 2025

Язык: Английский

Процитировано

0
Non-Invasive Delivery of CRISPR/Cas9 Ribonucleoproteins (Cas9 RNPs) into Cells via Nanoparticles for Membrane Transport DOI Creative Commons

Toshihiko Tashima

Pharmaceutics, Год журнала: 2025, Номер 17(2), С. 201 - 201

Опубликована: Фев. 6, 2025

The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system is a promising biotechnology tool for genome editing. However, in living organisms, several pharmacokinetic challenges arise, including off-target side effects due to incorrect distribution, low bioavailability caused by membrane impermeability, and instability resulting from enzymatic degradation. Therefore, innovative delivery strategies must be developed address these issues. Modified nanoparticles offer potential solution the non-invasive of CRISPR/Cas9 ribonucleoproteins (Cas9 RNPs). Cas9 RNPs encapsulated are protected degradation, similar how microRNAs shielded within exosomes. It well-established that certain materials, proteins, expressed selectively specific cell types. For example, α-7 nicotinic receptor endothelial neuronal cells, while αvβ3 integrin cancer cells. These endogenous materials can facilitate receptor-mediated endocytosis or transcytosis. Nanoparticles encapsulating coated with ligands targeting such receptors may internalized through mechanisms. Once internalized, could perform desired gene editing nucleus after escaping endosome mechanisms as proton sponge effect fusion. In this review, I discuss advantages delivering RNP-encapsulated

Язык: Английский

Процитировано

0
Integrin beta 1 and fibronectin mediate extracellular vesicle uptake and functional RNA delivery DOI Creative Commons

Omnia M. Elsharkasy,

Willemijn S. de Voogt,

Maria Laura Tognoli

и другие.

Journal of Biological Chemistry, Год журнала: 2025, Номер unknown, С. 108305 - 108305

Опубликована: Фев. 1, 2025

Extracellular vesicles (EVs) are cell-derived secreted by all cell types into the extracellular spaces. EVs comprise a heterogenous population of that carry bioactive molecules, such as proteins, lipids, and RNAs, which they can deliver to recipient cells. Over past few years, have been recognized for their vital role in intercellular communication, thereby various physiological pathological processes. In addition, increasingly being studied potential drug delivery vehicles. It is therefore crucial understand mechanisms molecular players underlying EV uptake functional cargo delivery. Several studies investigated pathways; nonetheless, governing transfer remain largely lacking. Here, we show, using CRISPR/Cas9-mediated reporter system, integrin β1 on cells plays an important EV-mediated RNA Additionally, both interference blocking antibodies, show association with α4 essential this process. We demonstrate α4β1 interacts through surface localized fibronectin via binding its Leucine-Aspartic acid-Valine (LDV) motif, interaction reduces Thus, identify key mechanism could potentially facilitate research biology pave way development novel therapeutic approaches targeting pathways lead

Язык: Английский

Процитировано

0
Lipo-Xenopeptide Polyplexes for CRISPR/Cas9 based Gene editing at ultra-low dose DOI Creative Commons

Janin Germer,

Anna-Lina Lessl,

Jana Pöhmerer

и другие.

Journal of Controlled Release, Год журнала: 2024, Номер 370, С. 239 - 255

Опубликована: Апрель 27, 2024

Double pH-responsive xenopeptide carriers containing succinoyl tetraethylene pentamine (Stp) and lipo amino fatty acids (LAFs) were evaluated for CRISPR/Cas9 based genome editing. Different carrier topologies, variation of LAF/Stp ratios LAF types as Cas9 mRNA/sgRNA polyplexes screened in three different reporter cell lines using genomic targets (Pcsk9, eGFP, mdx exon 23). One U-shaped bundle (B2)-shaped lipo-xenopeptides exhibiting remarkable efficiencies identified. Genome editing potency top observed at sub-nanomolar EC

Язык: Английский

Процитировано

3
A modular strategy for extracellular vesicle-mediated CRISPR-Cas9 delivery through aptamer-based loading and UV-activated cargo release DOI Open Access

Omnia M. Elsharkasy,

Charlotte V. Hegeman,

Ivana Lansweers

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Май 24, 2024

Abstract CRISPR-Cas9 gene editing technology offers the potential to permanently repair genes containing pathological mutations. However, efficient intracellular delivery of Cas9 ribonucleoprotein complex remains one major hurdles in its therapeutic application. Extracellular vesicles (EVs) are biological nanosized membrane released by cells, that play an important role intercellular communication. Due their innate capability transfer proteins, RNA, and various other cargos, EVs have emerged as a novel promising strategy for macromolecular biotherapeutics, including ribonucleoproteins. Here, we present versatile, modular loading Cas9. We leverage high affinity binding MS2 coat proteins (MCPs) fused EV-enriched aptamers incorporated into single guide RNAs (sgRNAs), combination with UV-activated photocleavable linker domain, PhoCl. Combined Vesicular stomatitis virus G (VSV-G) protein this platform enables subsequent complex, which shows critical dependence on incorporation activation domain. As approach does not require any direct fusion demonstrate can readily be exchanged variants, transcriptional activator dCas9-VPR adenine base editor ABE8e, confirmed sensitive fluorescent reporter assays. Taken together, describe robust successful delivery, applied CRISPR-Cas9-based genetic engineering well regulation, underlining EV-mediated strategies treatment diseases.

Язык: Английский

Процитировано

2
Lipid nanoparticle‐mediated RNA delivery for immune cell modulation DOI Creative Commons
Emily H. Kim,

Sridatta V. Teerdhala,

Marshall S. Padilla

и другие.

European Journal of Immunology, Год журнала: 2024, Номер 54(12)

Опубликована: Сен. 16, 2024

Lipid nanoparticles (LNPs) have emerged as the preeminent nonviral drug delivery vehicles for nucleic acid therapeutics, exemplified by their usage in mRNA COVID-19 vaccines. As a safe and highly modular platform, LNPs are attractive wide range of applications. In addition to vaccines, being utilized platforms other immunoengineering efforts, especially cancer immunotherapies modulating immune cells functionality via delivery. this review, we focus on methods applications LNP-based immunotherapy five cell types: T cells, NK macrophages, stem dendritic cells. Each these types has wide-reaching but comes with unique challenges barriers. By combining knowledge immunology nanotechnology, can be developed improved targeting transfection, ultimately working toward novel clinical therapeutics.

Язык: Английский

Процитировано

2
Targeted CRISPR/Cas9 Lipid Nanoparticles Elicits Therapeutic Genome Editing in Head and Neck Cancer DOI Creative Commons

Razan Masarwy,

Dan Peer,

Lior Stotsky‐Oterin

и другие.

Advanced Science, Год журнала: 2024, Номер unknown

Опубликована: Дек. 30, 2024

Squamous cell carcinomas of the head and neck (HNSCC) originate in upper aerodigestive tract, including oral cavity, pharynx, larynx. Current treatments locally advanced HNSCC often lead to high treatment failure, disease recurrence, resulting poor survival rates. Advances mRNA technologies lipid nanoparticle (LNP) delivery systems led several clinical trials involving LNP-CRISPR-Cas9 mRNA-based therapeutics. Despite these advances, achieving cell-type-specific extrahepatic is still challenging. This study introduces a safe effective intratumoral EGFR-targeted CRISPR-LNP strategy for knocking out SOX2, which cancer-specific gene. To assess their therapeutic potential, it shown that LNPs made from ionizable lipids with helper co-encapsulating Cas9 sgRNA targeting SOX2 (sgSOX2), ≈60% reduction viability vitro. Next, using xenograft mouse model, targeted 𝜶EGFR- CRISPR-sgSOX2-LNPs cells resulted 90% inhibition tumor growth increase > 84 days, disappearance observed 50% mice. These findings emphasize potential mRNA-Cas9-LNPs clinically accessible solid tumors, specifically reaching inducing persistent responses tumors high-recurrence rates like HNSCC.

Язык: Английский

Процитировано

2
Dual pH-responsive CRISPR/Cas9 ribonucleoprotein xenopeptide complexes for genome editing DOI Creative Commons

Xianjin Luo,

Janin Germer,

Tobias Burghardt

и другие.

European Journal of Pharmaceutical Sciences, Год журнала: 2024, Номер 205, С. 106983 - 106983

Опубликована: Дек. 7, 2024

Clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR associated (Cas) protein has been proved as a powerful tool for the treatment of genetic diseases. The Cas9 protein, when combined with single-guide RNA (sgRNA), forms Cas9/sgRNA ribonucleoprotein (RNP) capable targeting and editing genome. However, limited availability effective carriers restricted broader application CRISPR/Cas9 RNP. In this study, we evaluated dual pH-responsive amphiphilic xenopeptides (XPs) delivering These artificial lipo-XPs contain apolar cationizable lipoamino fatty acid (LAF) polar oligoaminoethylene units such succinoyl-tetraethylenepentamine (Stp) in various ratios U-shaped topologies. were screened functional RNP delivery four different reporter cell lines, including Duchenne muscular dystrophy (DMD) exon skipping model. Significantly enhanced cellular uptake into HeLa cells, endosomal disruption gal8-mRuby3 potent genome by several complexes was observed lines 5 nM sgRNA range. Comparing mRNA/sgRNA polyplexes DMD model demonstrated similar splice site high two molecular modalities. Based on these studies, analogues U1 LAF2-Stp LAF4-Stp2 structures deployed, tuning amphiphilicity Stp group replacement six oligoamino acids dmGtp, chGtp, dGtp, Htp, Stt, or GEIPA. most (containing chGtp GEIPA) further gene efficiency EC50 values 1 line. Notably, LAF2-dGtp reached 0.51 even upon serum incubation. Another carrier (LAF4-GEIPA2) complexing donor DNA, facilitated up to 43 % homology-directed repair (HDR) eGFPd2 cells visualized switch from green fluorescent (eGFP) blue (BFP). This study presents system tunable RNP/donor DNA polyplexes, offering an easily applicable strategy editing.

Язык: Английский

Процитировано

2