Transdermal delivery of CRISPR/Cas9-mediated melanoma gene therapy via polyamines-modified thermosensitive hydrogels

Abstract The main obstacles to the clinical application of the CRISPR/Cas9 system are off-target effects and low delivery efficiency. There is an urgent need to develop new delivery strategies and technologies. Three types of in situ injectable hydrogels with different electrical properties were cre...

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Main Authors: Meng Li, Songli Zhou, Suqin Zhang, Xingyu Xie, Junqi Nie, Qi Wang, Lixin Ma, Yibin Cheng, Jingwen Luo
Format: Article
Language:English
Published: BMC 2025-06-01
Series:Journal of Nanobiotechnology
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Online Access:https://doi.org/10.1186/s12951-025-03523-7
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author Meng Li
Songli Zhou
Suqin Zhang
Xingyu Xie
Junqi Nie
Qi Wang
Lixin Ma
Yibin Cheng
Jingwen Luo
author_facet Meng Li
Songli Zhou
Suqin Zhang
Xingyu Xie
Junqi Nie
Qi Wang
Lixin Ma
Yibin Cheng
Jingwen Luo
author_sort Meng Li
collection DOAJ
description Abstract The main obstacles to the clinical application of the CRISPR/Cas9 system are off-target effects and low delivery efficiency. There is an urgent need to develop new delivery strategies and technologies. Three types of in situ injectable hydrogels with different electrical properties were created to find the most secure and efficient sustained-release drug delivery system. After in vitro and in vivo comparisons, we found that the positively charged hydrogels had higher cellular uptake, stronger gene editing efficiency, greater cytotoxicity, longer tumor accumulation, and better anti-tumor efficacy than negatively charged and neutral hydrogels. We designed single guide RNA targeting the Y-box binding protein 1 (YB-1) gene and then used it to create a ribonucleoprotein complex with Cas9 protein. Doxorubicin was co-encapsulated into this positively charged hydrogel to create a co-delivery system. By knocking down YB-1, the expression of YB-1 was reduced, inhibiting the growth and migration of melanoma cells. The strategy of combining YB-1 gene editing and intratumoral injection enhanced the therapeutic effect of doxorubicin while reducing side effects.
format Article
id doaj-art-82de00689dda49d7a54519ab892d4981
institution DOAJ
issn 1477-3155
language English
publishDate 2025-06-01
publisher BMC
record_format Article
series Journal of Nanobiotechnology
spelling doaj-art-82de00689dda49d7a54519ab892d49812025-08-20T02:39:47ZengBMCJournal of Nanobiotechnology1477-31552025-06-0123112010.1186/s12951-025-03523-7Transdermal delivery of CRISPR/Cas9-mediated melanoma gene therapy via polyamines-modified thermosensitive hydrogelsMeng Li0Songli Zhou1Suqin Zhang2Xingyu Xie3Junqi Nie4Qi Wang5Lixin Ma6Yibin Cheng7Jingwen Luo8State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Province Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei UniversityState Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Province Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei UniversityState Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Province Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei UniversityState Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Province Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei UniversityState Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Province Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei UniversityDepartment of Thoracic Surgery, Tongji Hospital Affiliated to Tongji Medical College of Huazhong, University of Science & TechnologyState Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Province Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei UniversityState Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Province Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei UniversityState Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Province Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei UniversityAbstract The main obstacles to the clinical application of the CRISPR/Cas9 system are off-target effects and low delivery efficiency. There is an urgent need to develop new delivery strategies and technologies. Three types of in situ injectable hydrogels with different electrical properties were created to find the most secure and efficient sustained-release drug delivery system. After in vitro and in vivo comparisons, we found that the positively charged hydrogels had higher cellular uptake, stronger gene editing efficiency, greater cytotoxicity, longer tumor accumulation, and better anti-tumor efficacy than negatively charged and neutral hydrogels. We designed single guide RNA targeting the Y-box binding protein 1 (YB-1) gene and then used it to create a ribonucleoprotein complex with Cas9 protein. Doxorubicin was co-encapsulated into this positively charged hydrogel to create a co-delivery system. By knocking down YB-1, the expression of YB-1 was reduced, inhibiting the growth and migration of melanoma cells. The strategy of combining YB-1 gene editing and intratumoral injection enhanced the therapeutic effect of doxorubicin while reducing side effects.https://doi.org/10.1186/s12951-025-03523-7CRISPR/Cas9HydrogelPolyaminesIntratumoral injectionYB-1
spellingShingle Meng Li
Songli Zhou
Suqin Zhang
Xingyu Xie
Junqi Nie
Qi Wang
Lixin Ma
Yibin Cheng
Jingwen Luo
Transdermal delivery of CRISPR/Cas9-mediated melanoma gene therapy via polyamines-modified thermosensitive hydrogels
Journal of Nanobiotechnology
CRISPR/Cas9
Hydrogel
Polyamines
Intratumoral injection
YB-1
title Transdermal delivery of CRISPR/Cas9-mediated melanoma gene therapy via polyamines-modified thermosensitive hydrogels
title_full Transdermal delivery of CRISPR/Cas9-mediated melanoma gene therapy via polyamines-modified thermosensitive hydrogels
title_fullStr Transdermal delivery of CRISPR/Cas9-mediated melanoma gene therapy via polyamines-modified thermosensitive hydrogels
title_full_unstemmed Transdermal delivery of CRISPR/Cas9-mediated melanoma gene therapy via polyamines-modified thermosensitive hydrogels
title_short Transdermal delivery of CRISPR/Cas9-mediated melanoma gene therapy via polyamines-modified thermosensitive hydrogels
title_sort transdermal delivery of crispr cas9 mediated melanoma gene therapy via polyamines modified thermosensitive hydrogels
topic CRISPR/Cas9
Hydrogel
Polyamines
Intratumoral injection
YB-1
url https://doi.org/10.1186/s12951-025-03523-7
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