Methods in molecular biology (Clifton, N.J.)Sep 22, 2026

Using Tiny Fat Particles to Deliver CRISPR-Cas9 for Editing Genes

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗OA ↗

Updated Sep 23, 2026

Abstract

Lipid nanoparticles (LNPs) have been validated for the delivery of CRISPR components and have been used in multiple FDA-approved therapies.

LNPs are made from ionizable lipids, phospholipids, cholesterol, and PEG-lipids, allowing for efficient delivery of various therapeutic cargo.
They offer advantages over viral vectors, including lower immunogenicity and no genomic integration, making them suitable for genome editing applications.
Despite their benefits, LNPs face challenges such as limited tissue specificity and difficulties in escaping endosomes.
Recent improvements in lipid chemistry and surface modifications have enhanced their delivery performance.
Microfluidic mixing has been identified as a preferred method for LNP formulation due to its reproducibility and precise control over particle properties.

Simplified

Lipid nanoparticles (LNPs) are a clinically validated nonviral platform for the delivery of CRISPR-associated components. Composed of ionizable lipids, phospholipids, cholesterol, and PEG-lipids, LNPs enable the efficient encapsulation, protection, and cytosolic delivery of therapeutic cargo such as DNA, RNA, or proteins. The clinical relevance of LNPs has already been shown by multiple FDA-approved therapies, including siRNA-based treatments and mRNA vaccines. Compared with viral vectors, LNPs offer several advantages, including reduced immunogenicity, absence of genomic integration, scalable manufacturing, and flexibility in cargo size, while supporting transient expression, which is desirable for genome editing applications. However, challenges remain, including limited tissue specificity and inefficient endosomal escape. Recent advances in lipid chemistry optimization and surface modification have improved delivery performance. Among available formulation techniques, microfluidic mixing has emerged as a preferred method due to its reproducibility, scalability, and precise control over particle properties. This protocol describes a standardized microfluidic workflow for reproducible LNP formulation, providing practical guidance on lipid preparation, nanoparticle assembly, and quality control.

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Using Tiny Fat Particles to Deliver CRISPR-Cas9 for Editing Genes

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You found one interesting study. We’ll send the next 7.

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