Multifunctional Vector for Delivery of Genome Editing Plasmid Targeting β-Catenin to Remodulate Cancer Cell Properties.
Xiao-Yan HeBo-Ya LiuYan PengRen-Xi ZhuoSi-Xue ChengPublished in: ACS applied materials & interfaces (2018)
Accurate and efficient delivery of genome editing plasmids to targeted cells is of critical importance in genome editing. Herein, we prepared a multifunctional delivery vector with a combination of ligand-mediated selectivity and peptide-mediated transmembrane function to effectively deliver plasmids to targeted cancerous cells. In the delivery system, the clustered regularly interspaced short palindromic repeat-associated Cas9 nuclease (CRISPR-Cas9) plasmid is combined with protamine with membrane and nuclear translocating activities and co-precipitated with CaCO3, which is further decorated by AS1411-functionalized carboxymethyl chitosan and cell penetrating peptide (TAT)-functionalized carboxymethyl chitosan. The AS1411-mediated tumor cell/nuclear targeting and TAT-induced enhanced endocytosis result in obviously increased cellular uptake and nuclear transport. As a result, the CRISPR-Cas9 plasmid can be efficiently delivered to cancer cell nuclei to mediate genome editing, resulting in an efficacious knockout of CTNNB1 gene encoding β-catenin. More importantly, downregulation of β-catenin could effectively prevent its enrichment in nuclei and then significantly downregulate the expression of proteins, such as vimentin, Snail, MMP-2, MMP-9, CD44, Nanog, and Oct4 to prevent tumor progression and metastasis. The edited cancerous cells exhibit favorable remodulated properties including inhibited growth, suppressed migration and invasion, and reduced cancer stemness.
Keyphrases
- crispr cas
- genome editing
- induced apoptosis
- cancer therapy
- epithelial mesenchymal transition
- drug delivery
- cell cycle arrest
- cell proliferation
- escherichia coli
- poor prognosis
- single cell
- signaling pathway
- quantum dots
- stem cells
- endoplasmic reticulum stress
- cell death
- young adults
- genome wide
- papillary thyroid
- binding protein
- transcription factor
- molecularly imprinted
- squamous cell carcinoma
- dna methylation
- endothelial cells
- drug induced
- tandem mass spectrometry