Abstract
The epithelial-mesenchymal transition (EMT), a process in which epithelial cells undergo
a series of biochemical changes to acquire a mesenchymal phenotype, has been linked to tumor
metastasis. Here, we present a novel strategy for knocking out the EMT-related Cdh2 gene, which
encodes N-cadherin through CRISPR/Cas9-mediated gene editing by an ultrasound combined with
biosynthetic nanobubbles (Gas Vesicles, GVs). Polyethyleneimine were employed as a gene delivery
vector to deliver sgRNA into 4T1 cells that stably express the Cas9 protein, resulting in the stable
Cdh2 gene- knockout cell lines. TheWestern blotting assay confirmed the absence of an N-cadherin
protein in these Cdh2 gene-knockout 4T1 cell lines. Significantly reduced tumor cell migration was
observed in the Cdh2 gene-knockout 4T1 cells in comparison with the wild-type cells. Our study
demonstrated that an ultrasound combined with GVs could effectively mediate CRISPR/Cas9 gene
editing of a Cdh2 gene to inhibit tumor invasion and metastasis.