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Updated: Dec 14, 2025

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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CRISPR-Cas12a with an oAd Induces Precise and Cancer-Specific Genomic Reprogramming of EGFR and Efficient Tumor

A-Rum Yoon1, Bo-Kyeong Jung2, Eunyoung Choi3

  • 1Department of Bioengineering, College of Engineering, Hanyang University, Seoul 04763, Republic of Korea; Institute of Nano Science and Technology (INST), Hanyang University, Seoul 04763, Republic of Korea.

Molecular Therapy : the Journal of the American Society of Gene Therapy
|July 20, 2020
PubMed

Insights

Oncolytic adenovirus (oAd) delivers CRISPR-Cas12a genome editing tools specifically to cancer cells. This targeted approach disrupts oncogenes, halts tumor growth, and shows promise as a novel cancer therapy.

Area of Science:

  • Molecular Biology
  • Gene Editing
  • Oncology

Background:

  • CRISPR-Cas12a is a precise genome-editing endonuclease with therapeutic potential.
  • Oncolytic adenoviruses (oAds) offer cancer-specific delivery but haven't been used for genome editing.

Purpose of the Study:

  • To investigate the use of oAd for delivering CRISPR-Cas12a for cancer genome editing.
  • To evaluate the efficacy and specificity of oAd-mediated CRISPR-Cas12a in disrupting oncogenic pathways.

Main Methods:

  • Co-expression of Cas12a and a CRISPR RNA (crRNA) targeting the epidermal growth factor receptor (EGFR) gene within an oAd vector (oAd/Cas12a/crEGFR).
  • Intratumoral delivery of oAd/Cas12a/crEGFR into cancer models.
  • Assessment of EGFR gene editing, off-target activity, apoptosis induction, and tumor proliferation inhibition.

Main Results:

  • oAd/Cas12a/crEGFR achieved precise and efficient EGFR gene editing in a cancer-specific manner.
  • No detectable off-target nuclease activity was observed.
  • Significant antitumor effects, including apoptosis induction and proliferation inhibition, led to complete tumor regression in some mice.

Conclusions:

  • Single oAd vector-mediated CRISPR-Cas system enables precise genomic reprogramming.
  • This system demonstrates feasibility as an alternative cancer therapy with high specificity and potent antitumor activity.
Keywords:
CRISPRCas12aCpf1EGFRanti-tumor effectgenome editingoncolytic virusout-of-frame

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