Fast Creation of Single and Multiple Gene Knockouts in Human Cells Using CRISPR-Cas9

An improved method achieved high editing efficiency for generating multiple gene knockouts in HEK293T cells.

• The method utilizes sgRNA DNA strands and the CRISPR/Cas9 system for effective multi-gene knockouts.
• Target genes were selected from the apoptosis pathway, focusing on Exon 1 to create premature STOP codons.
• T7 endonuclease I assays confirmed the effectiveness of the sgRNAs for single and multiple gene knockouts.
• Deep sequencing analysis identified frameshift mutations and premature stop codons, supporting successful gene knockouts.
• The new approach may offer a more uniform and efficient editing profile than existing methods.

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