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Updated: Jan 6, 2026

Electroporation-Based CRISPR-Cas9-Mediated Gene Knockout in THP-1 Cells and Single-Cell Clone Isolation
Published on: February 28, 2025
Ku80 gene knockdown by the CRISPR/Cas9 technique affects the biological functions of human thyroid carcinoma cells
Yali Fan1, Jianying Li1, Wei Wei2
1Department of Respiratory Medicine, Xi'an Central Hospital, The Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710003, P.R. China.
Abstract:
In the present study, to evaluate the role of Ku80 in thyroid carcinoma (TC), 86 thyroid tissue samples from patients with a spectrum of thyroid disorders were examined for protein levels of Ku80, nuclear factor‑κB (NF‑κB) and RET/TC by immunohistochemistry. Furthermore, in TC cells, Ku80 mRNA was detected by reverse transcription‑quantitative PCR analysis and silenced using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‑associated protein 9 (Cas9) technique to assess its role. An antibody array was used to identify Ku80‑related regulatory genes. The protein levels of Ku80 in the TC tissues were significantly higher than those in non‑neoplastic adjacent tissue samples (P<0.01). The activation of NF‑kB and expression of RET/TC in the TC group were significantly increased (P<0.05) and were correlated with the protein expression of Ku80 (P<0.05). In papillary TC cells, the mRNA levels of Ku80 were high; Ku80 knockdown resulted in reductions in proliferation, invasion and colony formation, increased apoptosis, and reduced levels of proteins involved in MAPK signaling, cell proliferation and apoptosis. The high expression of Ku80 in TC was found to be associated with the expression of RET/TC and activation of NF‑κB, and Ku80 knockdown decreased the malignancy of TC cells.
Insights
Ku80 protein is elevated in thyroid carcinoma (TC) and drives cancer progression. Silencing Ku80 in TC cells reduced proliferation and invasion, suggesting Ku80 as a potential therapeutic target for thyroid cancer.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Thyroid carcinoma (TC) is a significant health concern.
- The role of Ku80 in the development and progression of TC requires further elucidation.
Purpose of the Study:
- To investigate the role of Ku80 in thyroid carcinoma.
- To assess the correlation between Ku80 expression and key cancer markers.
Main Methods:
- Immunohistochemistry on 86 thyroid tissue samples.
- Reverse transcription-quantitative PCR and CRISPR/Cas9 gene silencing in TC cells.
- Antibody array analysis to identify related genes.
Main Results:
- Ku80 protein levels were significantly higher in TC tissues compared to adjacent non-neoplastic tissues.
- Ku80 expression correlated with nuclear factor-κB (NF-κB) activation and RET/TC expression.
- Ku80 knockdown in papillary TC cells reduced proliferation, invasion, and colony formation, while increasing apoptosis.
Conclusions:
- Ku80 plays a crucial role in promoting thyroid carcinoma malignancy.
- Ku80 is associated with NF-κB activation and RET/TC expression in TC.
- Targeting Ku80 may offer a novel therapeutic strategy for thyroid cancer.

