Molecular therapy. Nucleic acidsJul 13, 2026

Precise gene editing creates liver cells with PNPLA3-I148M mutation to study fat-related cell damage

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Jul 14, 2026

Abstract

Clones with the G allele of the PNPLA3-I148M variant showed enhanced lipid storage and reduced sensitivity to palmitic acid-induced toxicity.

Lipid droplet accumulation was greater in clones with the G allele (GG, GC) compared to wild-type (CC) clones.
Cell viability assays indicated that G allele clones were less affected by palmitic acid-induced toxicity.
Distinct transcriptomic profiles were observed among different genotypes, suggesting varied regulation of lipid handling.
Heterozygous clones exhibited intermediate phenotypes, indicating allele-associated effects on lipotoxic responses.
The use of prime editing allowed for the generation of isogenic HepG2 clones to study genetic variant impacts.

Simplified

The PNPLA3-I148M (rs738409 C>G) variant is a well-established genetic determinant of hepatic steatosis, fibrosis, and hepatocellular carcinoma. However, its precise functional impact on hepatocyte physiology remains incompletely understood. Here, we employed a GFP-linked all-in-one prime editing system to generate isogenic HepG2 clones representing all PNPLA3 genotypes in an identical genetic background. Following palmitic acid (PA) treatment, we evaluated genotype-specific responses to lipotoxic stress. Lipid droplet accumulation and cell viability assays showed that clones harboring the G allele (GG, GC) exhibited enhanced lipid storage and reduced sensitivity to PA-induced toxicity compared to wild-type (CC) clones. Transcriptomic profiling revealed distinct expression patterns among genotypes, consistent with differential regulation of lipid handling and stress adaptation programs. Heterozygous clones displayed partially intermediate phenotypes, suggesting allele-associated effects of PNPLA3-I148M on hepatocellular lipotoxic responses. This study demonstrates that prime editing enables a precise approach for modeling disease-associated genetic variants in human hepatocytes and provides a platform to investigate genotype-associated cellular responses. By combining precise genome editing with human cell modeling, this study establishes a useful system for dissecting PNPLA3-I148M-related genotype-phenotype relationships and for future mechanistic studies of genetically defined liver disease phenotypes.

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PubMed

Precise gene editing creates liver cells with PNPLA3-I148M mutation to study fat-related cell damage

Abstract

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