gene editing

Supporting data for "Uncovering cellular response to ionizable lipid-induced endolysosomal damage with multimodal imaging"

Lipid nanoparticles (LNPs) are the leading non-viral platform for nucleic acid delivery, validated clinically by their use in mRNA vaccines and the siRNA drug patisiran. Nevertheless, their wider use in protein-replacement and gene-editing therapeutics need repeated doses is limited by inflammatory side effects caused...

Production of Lentiviral Vectors Encoding the CRISPR-Cas13d System for RNA Targeting.

A detailed protocol for the production of high-titer, third-generation LVs with a dual-expression cassette for Cas13d and a customizable guide RNA is described, which can be used for stable transduction and efficient RNA knockdown in a broad range of mammalian cell types.

Adeno-Associated Viral Vector (AAV)-Mediated In Vivo CRISPR-Cas9 Delivery.

This chapter detail a methodology using a rAAV vector carrying Staphylococcus aureus Cas9 (SaCas9) to target the murine Hao1 gene in liver parenchymal cells as a curative treatment for primary hyperoxaluria type 1 (PH1).

Establishment of a high-efficiency hairy root transformation system for Forsythia suspensa and enhancement of transformation efficiency by developmental regulators

Forsythia suspensa (Thunb.) Vahl is an economically important woody medicinal plant, yet the lack of efficient genetic transformation protocols hinders genomic and molecular breeding studies. Conventional Agrobacterium tumefaciens -mediated transformation faces low efficiency and long culture cycles, while Agrobacteriu...

Emerging insights and breakthroughs in genetic advancement and phytopathology of Sclerotinia stem rot in rapeseed-mustard

Production and Storage of High-Capacity Adenoviral Vectors for the Delivery of Advanced CRISPR Systems.

Genome editing based on engineered CRISPR systems is advancing rapidly, with the field increasingly moving toward approaches that avoid the induction of mutagenic double-stranded DNA breaks (e.g., RNA-programmable base editing, prime editing, and donor DNA transposition). These nuclease-free strategies often rely on la...

Nucleofection-Based CRISPR/Cas Delivery in Human T Cells for Immunotherapy Applications.

Electroporation-based delivery of CRISPR/Cas systems has emerged as a powerful and versatile approach for gene editing in primary human T cells, enabling efficient, transient, and nonviral modification while minimizing genomic integration risks. This chapter focuses on the principles and practical implementation of ele...

Quantitative Analysis of CRISPR Encoding Plasmid DNA–Nanocarrier Interactions by Gel Electrophoresis and Densitometry

Nanoparticle-based carriers are key systems for the transport and delivery of nucleic acids in gene therapy, RNA therapeutics, and genome editing applications. The development of efficient delivery systems for CRISPR–Cas technologies requires reliable methods to evaluate nanocarrier–nucleic acid interactions, binding e...

CRISPR/Cas Systems: Biological Basis and Genome Editing Applications.

Clustered regularly interspaced short palindromic repeats (CRISPR) and associated (Cas) systems have revolutionized the field of genome engineering by providing versatile, efficient, and programmable tools for precise genetic manipulation. Originally identified as an adaptive immune mechanism in prokaryotes, CRISPR/Cas...

Beyond conventional immunosuppression: tailoring perioperative management protocols for clinical xenotransplantation

This review proposes a standardized perioperative management protocol that is applicable to clinical xenotransplantation and addresses consumptive coagulopathy; targeted suppression of macrophage- and neutrophil-driven innate inflammatory cascades, alongside activation of xenoreactive natural killer cells.

Systematic Profiling of ssODN Determinants Establishes Optimized LbCas12a-Mediated HDR for Precise Genome Editing in Flammulina filiformis

The results not only establish a versatile precision-editing platform for F. filiformis through optimized ssODN design, but also offer tentative and transferable insights into the HDR-mediated genome editing of other heterokaryotic basidiomycetes facing similar genetic operation challenges.

Overview of Delivery Methods for Gene Editing.

This chapter emphasizes that the strategic selection of a delivery vehicle must be harmonized with the specific cargo and the unique biological requirements of the target tissue to ensure therapeutic success.