Cascade dynamic assembly/disassembly of DNA nanoframework enabling the controlled delivery of CRISPR-Cas9 system.
Nachuan SongYiwen ChuShuai LiYuhang DongXiaoting FanJianpu TangYanfei GuoGuangshuai TengChi YaoDayong YangPublished in: Science advances (2023)
CRISPR-Cas9 has been explored as a therapeutic agent for down-regulating target genes; the controlled delivery of Cas9 ribonucleoprotein (RNP) is essential for therapeutic efficacy and remains a challenge. Here, we report cascade dynamic assembly/disassembly of DNA nanoframework (NF) that enables the controlled delivery of Cas9 RNP. NF was prepared with acrylamide-modified DNA that initiated cascade hybridization chain reaction (HCR). Through an HCR, single-guide RNA was incorporated to NF; simultaneously, the internal space of NF was expanded, facilitating the loading of Cas9 protein. NF was designed with hydrophilic acylamino and hydrophobic isopropyl, allowing dynamic swelling and aggregation. The responsive release of Cas9 RNP was realized by introducing disulfide bond-containing N , N -bis(acryloyl)cystamine that was specifically in response to glutathione of cancer cells, triggering the complete disassembly of NF. In vitro and in vivo investigations demonstrated the high gene editing efficiency in cancer cells, the hypotoxicity in normal cells, and notable antitumor efficacy in a breast cancer mouse model.
Keyphrases
- crispr cas
- genome editing
- signaling pathway
- lps induced
- pi k akt
- nuclear factor
- oxidative stress
- circulating tumor
- induced apoptosis
- single molecule
- mouse model
- cell free
- inflammatory response
- ionic liquid
- nucleic acid
- dna methylation
- small molecule
- young adults
- cell proliferation
- cell death
- drug delivery
- cancer therapy
- endoplasmic reticulum stress
- protein protein