CRISPR (clustered regularly interspaced short palindromic repeats) genome editing holds promise in the treatment of genetic diseases that currently lack effective long-term therapies. Patients with alpha-1 antitrypsin (AAT) deficiency develop progressive lung disease due to the loss of AAT's antiprotease function and liver disease due to a toxic gain of function of the common mutant allele. However, it remains unknown whether CRISPR-mediated AAT correction in the liver, where AAT is primarily expressed, can correct either or both defects. Here we show that AAV delivery of CRISPR can effectively correct Z-AAT mutation in the liver of a transgenic mouse model. Specifically, we co-injected two AAVs: one expressing Cas9 and another encoding an AAT guide RNA and homology-directed repair template. In both neonatal and adult mice, this treatment partially restored M-AAT in the serum. Furthermore, deep sequencing confirmed both indel mutations and precise gene correction in the liver, permitting careful analysis of gene editing events in vivo. This study demonstrates a proof of concept for the application of CRISPR-Cas9 technology to correct AAT mutations in vivo and validates continued exploration of this approach for the treatment of patients with AAT deficiency.
In Vivo Genome Editing Partially Restores Alpha1-Antitrypsin in a Murine Model of AAT Deficiency
Chun‐Qing Song,Dan Wang,Tingting Jiang,K. O'Connor,Qiushi Tang,Lingling Cai,Xiangrui Li,Z. Weng,Hao Yin,G. Gao,C. Mueller,T. Flotte,Wen Xue
Published 2018 in Human Gene Therapy
ABSTRACT
PUBLICATION RECORD
- Publication year
2018
- Venue
Human Gene Therapy
- Publication date
2018-08-01
- Fields of study
Biology, Medicine
- Identifiers
- External record
- Source metadata
Semantic Scholar, PubMed
CITATION MAP
EXTRACTION MAP
CLAIMS
CONCEPTS
- adeno-associated virus delivery
Use of adeno-associated virus vectors to deliver the editing components into mouse liver cells.
Aliases: AAV delivery, AAVs
- adult mice
Mature mice treated to test whether editing also works after development.
Aliases: mature mice
- alpha-1 antitrypsin deficiency
A genetic condition in which defective alpha-1 antitrypsin production leads to lung and liver disease.
Aliases: AAT deficiency
- crispr-cas9 genome editing
A genome editing approach that uses the Cas9 nuclease and guide RNA targeting to modify DNA at a chosen locus.
Aliases: CRISPR, CRISPR-Cas9
- deep sequencing
High-throughput sequencing used to characterize editing outcomes in the liver DNA.
Aliases: sequencing
- homology-directed repair template
A donor DNA template supplied with the editing system to support precise sequence correction after Cas9 cutting.
Aliases: HDR template
- m-aat
The mouse alpha-1 antitrypsin protein measured in serum as the readout of restored expression.
Aliases: mouse AAT
- neonatal mice
Young mice treated at the neonatal stage to test editing in early life.
Aliases: newborn mice
- transgenic mouse model
A genetically engineered mouse model carrying the alpha-1 antitrypsin disease allele used for in vivo editing experiments.
Aliases: murine model
- z-aat mutation
The common mutant alpha-1 antitrypsin allele targeted for correction in the liver.
Aliases: Z-AAT allele, mutant allele
REFERENCES
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CITED BY
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