|
| Status |
Public on Nov 10, 2023 |
| Title |
Efficient and Safe Therapeutic Use of Paired Cas9-Nickases for Primary Hyperoxaluria Type 1 |
| Organism |
Mus musculus |
| Experiment type |
Other
|
| Summary |
The therapeutic use of adeno-associated viral vector (AAV)-mediated gene disruption using CRISPR-Cas9 is limited by potential off-target modifications and the risk of uncontrolled integration of vector genomes into CRISPR-mediated double-strand breaks. To address these concerns, we explored the use of AAV-delivered paired Staphylococcus aureus nickases (D10ASaCas9) to target the Hao1 gene for the treatment of primary hyperoxaluria type 1 (PH1). Our study demonstrated effective Hao1 gene disruption, a significant decrease in glycolate oxidase expression, and a therapeutic effect in PH1 mice. The assessment of undesired genetic modifications through CIRCLE-seq and CAST-Seq analyses revealed neither off-target activity nor chromosomal translocations. Importantly, the use of paired-D10ASaCas9 resulted in a significant reduction in AAV integration at the target site compared to SaCas9 nuclease. Additionally, our study highlights the limitations of current analytical tools in characterizing modifications introduced by paired D10ASaCas9, necessitating the development of a custom pipeline for more accurate characterization. These results describe a positive advance towards a safe and effective potential long-term treatment for PH1 patients.
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| Overall design |
Mouse embryonic fibroblasts (MEFs) were treated with single and paired SaCas9 nucleases targeting Hao1. Genotoxicity was assesed via CAST-Seq.
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| Contributor(s) |
Andrieux G, Torella L, Gonzalez-Aseguinolaza G, Klermund J, Cathomen T |
| Citation(s) |
38182795 |
| |
| Submission date |
Oct 19, 2023 |
| Last update date |
Feb 09, 2024 |
| Contact name |
Geoffroy Andrieux |
| Organization name |
University clinics Freiburg
|
| Street address |
Breisacherstr 153
|
| City |
Freiburg |
| ZIP/Postal code |
79110 |
| Country |
Germany |
| |
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| Platforms (1) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
|
| Samples (14)
|
| GSM7849065 |
Untreated sample used as control replicate1 |
| GSM7849066 |
Untreated sample used as control replicate2 |
| GSM7849067 |
Untreated sample used as control replicate3 |
| GSM7849068 |
Untreated sample used as control replicate4 |
| GSM7849069 |
CAST-seq on samples treated with SaCas9 paired with g1 replicate1 |
| GSM7849070 |
CAST-seq on samples treated with SaCas9 paired with g1 replicate2 |
| GSM7849071 |
CAST-seq on samples treated with SaCas9 paired with g2 replicate1 |
| GSM7849072 |
CAST-seq on samples treated with SaCas9 paired with g2 replicate2 |
| GSM7849073 |
CAST-seq on samples treated with SaCas9 paired with g2 replicate3 |
| GSM7849074 |
CAST-seq on samples treated with SaCas9 paired with g2 replicate4 |
| GSM7849075 |
CAST-seq on samples treated with SaCas9 paired with g1+g2 (paired nuclease) replicate1 |
| GSM7849076 |
CAST-seq on samples treated with SaCas9 paired with g1+g2 (paired nuclease) replicate2 |
| GSM7849077 |
CAST-seq on samples treated with SaCas9 paired with g1+g2 (paired nuclease) replicate3 |
| GSM7849078 |
CAST-seq on samples treated with SaCas9 paired with g1+g2 (paired nuclease) replicate4 |
|
| Relations |
| BioProject |
PRJNA1029956 |
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