Single-AAV Delivery of the SpCas9 Nickase Improves CAG/CTG Repeat contraction in a mouse model of Huntington's disease
- 1Dementia Research Institute at Cardiff University
Abstract
The best studied Cas9 enzyme, from Streptococcus pyogenes, is thought to be too large to be efficiently packaged into a single adeno-associated virus (AAV) together with a single-guide (sg) RNA cassette, requiring a dual-vector system. Here we have circumvented this limitation and packaged the full CRISPR system into a single AAV by using a single promoter driving the expression of both the Cas9 enzyme and a sgRNA. We compared the ability of the dual and single vector systems to induce contraction of CAG/CTG repeats in the striatum of a Huntington's disease model. We show that the all-in-one system led to larger contractions of the CAG/CTG repeats. This approach provides a more efficient and less costly approach for in vivo gene editing and improves the translational potential of AAV-mediated delivery of Cas9.
Related
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BibTeX
@misc{murillo2026singleaav,
title = {{Single-AAV Delivery of the SpCas9 Nickase Improves CAG/CTG Repeat contraction in a mouse model of Huntington's disease}},
author = {A. Murillo and A. N. Aston and M. Lelos and V. Dion},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.30.762787},
url = {https://www.biorxiv.org/content/10.64898/2026.10.30.762787v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Single-AAV Delivery of the SpCas9 Nickase Improves CAG/CTG Repeat contraction in a mouse model of Huntington's disease
AU - A. Murillo
AU - A. N. Aston
AU - M. Lelos
AU - V. Dion
PY - 2026
DA - 2026/10/30
PB - bioRxiv
DO - 10.64898/2026.10.30.762787
UR - https://www.biorxiv.org/content/10.64898/2026.10.30.762787v1
AB - The best studied Cas9 enzyme, from Streptococcus pyogenes, is thought to be too large to be efficiently packaged into a single adeno-associated virus (AAV) together with a single-guide (sg) RNA cassette, requiring a dual-vector system. Here we have circumvented this limitation and packaged the full CRISPR system into a single AAV by using a single promoter driving the expression of both the Cas9 enzyme and a sgRNA. We compared the ability of the dual and single vector systems to induce contraction of CAG/CTG repeats in the striatum of a Huntington's disease model. We show that the all-in-one system led to larger contractions of the CAG/CTG repeats. This approach provides a more efficient and less costly approach for in vivo gene editing and improves the translational potential of AAV-mediated delivery of Cas9.
ER -