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Co-delivery of Cas9 mRNA and guide RNAs for editing of LGMN gene represses breast cancer cell metastasis

Sci Rep. 2024-04; 
Yue Wang, Yatu Peng, Guanghui Zi, Jin Chen, Baowei Peng
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Proteins, Expression, Isolation and Analysis … The extracted protein was incubated with specific primary antibodies to spCas9 (GenScript, A01935-40, 1:500) and tubulin (ORIGENE, F001, 1:3000). Secondary antibody HRP-Donkey-… Get A Quote

摘要

Legumain (or asparagine endopeptidase/AEP) is a lysosomal cysteine endopeptidase associated with increased invasive and migratory behavior in a variety of cancers. In this study, co-delivery of Cas9 mRNA and guide RNA (gRNA) by lipid nanoparticles (LNP) for editing of LGMN gene was performed. For in-vitro transcription (IVT) of gRNA, two templates were designed: linearized pUC57-T7-gRNA and T7-gRNA oligos, and the effectiveness of gRNA was verified in multiple ways. Cas9 plasmid was modified and optimized for IVT of Cas9 mRNA. The effects of LGMN gene editing on lysosomal/autophagic function and cancer cell metastasis were investigated. Co-delivery of Cas9 mRNA and gRNA resulted in impaired lysosomal/autophagic... More

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