Nanocarriers for intracellular delivery of proteins in biomedical applications: strategies and recent advances DOI Creative Commons
Chuanda Zhu, Jing Mu, Ling Liang

et al.

Journal of Nanobiotechnology, Journal Year: 2024, Volume and Issue: 22(1)

Published: Nov. 10, 2024

Protein drugs are of great importance in maintaining the normal functioning living organisms. Indeed, they have been instrumental combating tumors and genetic diseases for decades. Among these pharmaceutical agents, those that target intracellular components necessitate use therapeutic proteins to exert their effects within targeted cells. However, protein is limited by short half-life potential adverse physiological environment. The advent nanoparticles offers a promising avenue prolonging drugs. This achieved encapsulating proteins, thereby safeguarding biological activity ensuring precise delivery into nanomaterial-based drug system mitigates rapid hydrolysis unwarranted diffusion minimizing side circumventing limitations inherent traditional techniques like electroporation. review examines established systems, including based on polymers, liposomes, nanoparticles. We delve operational principles transport mechanisms nanocarriers, discussing various considerations essential designing cutting-edge platforms. Additionally, we investigate innovative designs applications cytosolic systems medical research clinical practice, particularly areas tumor treatment, gene editing fluorescence imaging. sheds light current restrictions anticipates future avenues, aiming foster continued advancement this field.

Language: Английский

Strategies and mechanisms for endosomal escape of therapeutic nucleic acids DOI Creative Commons

Melina Grau,

Ernst Wagner

Current Opinion in Chemical Biology, Journal Year: 2024, Volume and Issue: 81, P. 102506 - 102506

Published: Aug. 1, 2024

Despite impressive recent establishment of therapeutic nucleic acids as drugs and vaccines, their broader medical use is impaired by modest performance in intracellular delivery. Inefficient endosomal escape presents a major limitation responsible for inadequate cytosolic cargo release. Depending on the carrier, this barrier can strongly limit or even abolish acid Different strategies hypothesized mechanisms are reviewed.

Language: Английский

Citations

10
CRISPR/Cas9 Ribonucleoprotein Delivery Enhanced by Lipo-Xenopeptide Carriers and Homology-Directed Repair Modulators: Insights from Reporter Cell Lines DOI Open Access

Xianjin Luo,

Eric Weidinger,

Tobias Burghardt

et al.

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(9), P. 4361 - 4361

Published: May 3, 2025

CRISPR-Cas9 genome editing is a versatile platform for studying and treating various diseases. Homology-directed repair (HDR) with DNA donor templates serves as the primary pathway gene correction in therapeutic applications, but its efficiency remains significant challenge. This study investigates strategies to enhance using T-shaped lipo-xenopeptide (XP)-based Cas9 RNP/ssDNA delivery system combined HDR enhancers. Nu7441, known DNA-PKcs inhibitor, was found be most effective enhancing HDR-mediated correction. An over 10-fold increase achieved by Nu7441 HeLa-eGFPd2 cells, peak of 53% at 5 nM RNP concentration up 61% confirmed Sanger sequencing. Surprisingly, total including non-homologous end joining (NHEJ) also improved. For example, boosted exon skipping via NHEJ-mediated splice site destruction 30-fold DMD reporter cell model. modulated cycle reducing cells G1 phase extending S G2/M phases without compromising cellular uptake or endosomal escape. The enhancement widely applicable across several lines, carriers (LAF-XPs), mRNA/sgRNA/ssDNA polyplexes. These findings highlight novel counterintuitive role an enhancer both efficiency, presenting promising strategy RNP-based therapy.

Language: Английский

Citations

0
Dual pH-responsive CRISPR/Cas9 ribonucleoprotein xenopeptide complexes for genome editing DOI Creative Commons

Xianjin Luo,

Janin Germer,

Tobias Burghardt

et al.

European Journal of Pharmaceutical Sciences, Journal Year: 2024, Volume and Issue: 205, P. 106983 - 106983

Published: Dec. 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.

Language: Английский

Citations

2
Nanocarriers for intracellular delivery of proteins in biomedical applications: strategies and recent advances DOI Creative Commons
Chuanda Zhu, Jing Mu, Ling Liang

et al.

Journal of Nanobiotechnology, Journal Year: 2024, Volume and Issue: 22(1)

Published: Nov. 10, 2024

Protein drugs are of great importance in maintaining the normal functioning living organisms. Indeed, they have been instrumental combating tumors and genetic diseases for decades. Among these pharmaceutical agents, those that target intracellular components necessitate use therapeutic proteins to exert their effects within targeted cells. However, protein is limited by short half-life potential adverse physiological environment. The advent nanoparticles offers a promising avenue prolonging drugs. This achieved encapsulating proteins, thereby safeguarding biological activity ensuring precise delivery into nanomaterial-based drug system mitigates rapid hydrolysis unwarranted diffusion minimizing side circumventing limitations inherent traditional techniques like electroporation. review examines established systems, including based on polymers, liposomes, nanoparticles. We delve operational principles transport mechanisms nanocarriers, discussing various considerations essential designing cutting-edge platforms. Additionally, we investigate innovative designs applications cytosolic systems medical research clinical practice, particularly areas tumor treatment, gene editing fluorescence imaging. sheds light current restrictions anticipates future avenues, aiming foster continued advancement this field.

Language: Английский

Citations

1