CRISPR/Cas9 Gene Editing System Can Alter Gene Expression and Induce DNA Damage Accumulation

Area of Science:

• Molecular Biology
• Genomics
• Biotechnology

Background:

• Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas) systems are powerful gene editing tools.
• Previous research indicates CRISPR/Cas gene editing can lead to P53 activation, large deletions, and chromosomal variations.

Purpose of the Study:

• To investigate the impact of CRISPR/Cas9 gene editing on host cell gene expression.
• To assess potential safety risks and cancer-related changes induced by CRISPR/Cas9 gene editing.

Main Methods:

• Transcriptome sequencing was employed to detect gene expression changes in host cells post-CRISPR/Cas9 editing.
• Gene ontology and KEGG pathway enrichment analyses were performed.
• Analysis included assessment of gene editing efficiency, alternative splicing, fusion gene formation, and DNA damage response activation.

Main Results:

• CRISPR/Cas9 gene editing significantly altered host cell gene expression, with the number of differentially expressed genes correlating with editing efficiency.
• Alternative splicing was observed at random sites, and single-site editing did not consistently produce fusion genes.
• While cell growth was not affected, the DNA damage response protein γH2AX was activated, indicating cellular stress.

Conclusions:

• CRISPR/Cas9 gene editing can induce significant alterations in gene expression and activate DNA damage responses.
• The observed changes suggest potential cancer-related implications, necessitating further research into the safety of CRISPR/Cas9 systems.
• This study provides crucial data for understanding and mitigating the risks associated with CRISPR/Cas9 gene editing applications.