Efficient shRNA-based knockdown of multiple target genes for cell therapy using a chimeric miRNA cluster platform.

ABSTRACT: Genome engineering technologies are powerful tools in cell-based immunotherapy to optimize or fine-tune cell functionalities. However, their use for multiple gene edits poses relevant biological and technical challenges. Short hairpin RNA (shRNA)-based cell engineering bypasses these criticalities and represents a valid alternative to CRISPR-based gene editing. Here, we describe a microRNA (miRNA)-based multiplex shRNA platform obtained by combining highly efficient miRNA scaffolds into a chimeric cluster, to deliver up to four shRNA-like sequences. Thanks to its limited size, our cassette could be deployed in a one-step process along with all the CAR components, streamlining the generation of engineered CAR T cells. The plug-and-play design of the shRNA platform allowed us to swap each sh

SUBMITTER: Rossi M 

PROVIDER: S-EPMC10548280 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature