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Updated: May 3, 2026

Electroporation-Based CRISPR-Cas9-Mediated Gene Knockout in THP-1 Cells and Single-Cell Clone Isolation
Published on: February 28, 2025
Knockin' on pHeaven's door: a fast and reliable high-throughput compatible zero-background cloning procedure
Quirino Schefer1, Sandy Hallmann, Carsten Grötzinger
1Department of Hepatology and Gastroenterology, Molecular Cancer Research Center (MKFZ), Charité - Universitätsmedizin Berlin, Campus Virchow-Klinikum, 13353, Berlin, Germany.
Abstract:
We created a pair of vectors allowing simple and efficient molecular cloning of any gene of interest with minimal consumption of time, labor and material. This system is applicable for standard molecular cloning, high-throughput cloning and generation of fusion protein libraries as well as for more complex gene assembly purposes. Also, this zero-background procedure allows going from cDNA to gene expression analysis in a defined vector in <2 days.
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