التفاصيل البيبلوغرافية
| العنوان: |
In vivo prime editing of a metabolic liver disease in mice. |
| المؤلفون: |
Böck, Desirée, Rothgangl, Tanja, Villiger, Lukas, Schmidheini, Lukas, Matsushita, Mai, Mathis, Nicolas, Ioannidi, Eleonora, Rimann, Nicole, Grisch-Chan, Hiu Man, Kreutzer, Susanne, Kontarakis, Zacharias, Kopf, Manfred, Thöny, Beat, Schwank, Gerald |
| المصدر: |
Science Translational Medicine; 3/16/2022, Vol. 14 Issue 636, p1-12, 12p |
| مصطلحات موضوعية: |
GENOME editing, LIVER diseases, DOUBLE-strand DNA breaks, METABOLIC disorders, GENETIC disorders, HEPATITIS |
| مستخلص: |
Prime editing is a highly versatile CRISPR-based genome editing technology that works without DNA double-strand break formation. Despite rapid technological advances, in vivo application for the treatment of genetic diseases remains challenging. Here, we developed a size-reduced SpCas9 prime editor (PE) lacking the RNaseH domain (PE2ΔRnH) and an intein-split construct (PE2 p.1153) for adeno-associated virus–mediated delivery into the liver. Editing efficiencies reached 15% at the Dnmt1 locus and were further elevated to 58% by delivering unsplit PE2ΔRnH via human adenoviral vector 5 (AdV). To provide proof of concept for correcting a genetic liver disease, we used the AdV approach for repairing the disease-causing Pahenu2 mutation in a mouse model of phenylketonuria (PKU) via prime editing. Average correction efficiencies of 11.1% (up to 17.4%) in neonates led to therapeutic reduction of blood phenylalanine, without inducing detectable off-target mutations or prolonged liver inflammation. Although the current in vivo prime editing approach for PKU has limitations for clinical application due to the requirement of high vector doses (7 × 1014 vg/kg) and the induction of immune responses to the vector and the PE, further development of the technology may lead to curative therapies for PKU and other genetic liver diseases. Preclinical prime editing for phenylketonuria: Prime editing nicks only one strand of DNA and allows for more versatile and precise editing than CRISPR-based strategies but is still challenging to apply in vivo. To fit a prime editor (PE) into commonly used gene editing vectors, Böck et al. developed a smaller yet still effective SpCas9 PE lacking its RNaseH domain. Adenoviral delivery of the PE ameliorated excess blood phenylalanine in a phenylalanine hydroxylase (Pah)–mutated neonate mouse model of phenylketonuria. Immune responses were however induced in response to the vector and PE, indicating further challenges remain for clinical translation. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
