Promoterless gene targeting without nucleases rescues lethality of a Crigler-Najjar syndrome mouse model

Fabiola Porro, Giulia Bortolussi, Adi Barzel, Alessia De Caneva, Alessandra Iaconcig, Simone Vodret, Lorena Zentilin, Mark A. Kay, Andrés F. Muro*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Crigler-Najjar syndrome type I (CNSI) is a rare monogenic disease characterized by severe neonatal unconjugated hyperbilirubinemia with a lifelong risk of neurological damage and death. Liver transplantation is the only curative option, which has several limitations and risks. We applied an in vivo gene targeting approach based on the insertion, without the use of nucleases, of a promoterless therapeutic cDNA into the albumin locus of a mouse model reproducing all major features of CNSI. Neonatal transduction with the donor vector resulted in the complete rescue from neonatal lethality, with a therapeutic reduction in plasma bilirubin lasting for at least 12 months, the latest time point analyzed. Mutant mice, which expressed about 5–6% of WT Ugt1a1 levels, showed normal liver histology and motor-coordination abilities, suggesting no functional liver or brain abnormalities. These results proved that the promoterless gene therapy is applicable for CNSI, providing therapeutic levels of an intracellular ER membrane-bound enzyme responsible for a lethal liver metabolic disease.

Original languageEnglish
Pages (from-to)1346-1355
Number of pages10
JournalEMBO Molecular Medicine
Volume9
Issue number10
DOIs
StatePublished - Oct 2017

Funding

FundersFunder number
Department of Life Sciences
National Institutes of Health
National Heart, Lung, and Blood InstituteR01HL064274
Università degli Studi di Trieste

    Keywords

    • brain damage
    • genetic liver disease
    • homologous recombination
    • hyperbilirubinemia
    • kernicterus

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