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Published on: April 10, 2018
CRISPR-Cas9-Mediated Silencing of CD44 in Human Highly Metastatic Osteosarcoma Cells
Background/Aims:
Metastasis is the major cause of death in patients with osteosarcoma. There is an urgent need to identify molecular markers that promote metastasis. Cluster of differentiation 44 is a receptor for hyaluronic acid (HA) and HA-binding has been proven to participate in various biological tumor activities, including tumor progression and metastasis.
Methods:
We performed a meta-analysis to investigate the relationship between CD44 expression, survival, and metastasis in patients with osteosarcoma. We then utilized the CRISPR-Cas9 system to specifically silence CD44 in highly metastatic human osteosarcoma cells (MNNG/HOS and 143B) and further determined the functional effects of CD44 knockout in these cells.
Results:
The meta-analysis demonstrated that a high level of CD44 may predict poor survival and higher potential of metastasis in patients with osteosarcoma. The expression of CD44 in highly metastatic human osteosarcoma cell lines was efficiently blocked by CRISPR-Cas9. When CD44 was silenced, the proliferation and spheroid formation of these osteosarcoma cells was inhibited under 3-D culture conditions. Furthermore, the migratory and invasive functions were also impaired in these highly metastatic osteosarcoma cells.
Conclusion:
These results suggest that developing new strategies to target CD44 in osteosarcoma may prevent metastasis and improve the clinical outcome of osteosarcoma patients.
Insights
High CD44 expression in osteosarcoma correlates with poor survival and increased metastasis. Silencing CD44 inhibited cancer cell proliferation and migration, suggesting CD44 as a therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Metastasis is a primary cause of mortality in osteosarcoma patients.
- Identifying molecular drivers of osteosarcoma metastasis is crucial.
- Cluster of differentiation 44 (CD44) is implicated in tumor progression and metastasis.
Purpose of the Study:
- To investigate the association between CD44 expression and osteosarcoma patient outcomes.
- To evaluate the functional role of CD44 in osteosarcoma metastasis using CRISPR-Cas9 gene editing.
Main Methods:
- A meta-analysis was conducted to examine CD44 expression, survival, and metastasis in osteosarcoma.
- CRISPR-Cas9 was used to silence CD44 in highly metastatic osteosarcoma cell lines.
- Functional assays assessed the impact of CD44 knockout on cell proliferation, spheroid formation, migration, and invasion.
Main Results:
- Meta-analysis indicated high CD44 levels predict poorer survival and increased metastasis risk.
- CRISPR-Cas9 efficiently blocked CD44 expression in osteosarcoma cells.
- CD44 silencing inhibited proliferation, spheroid formation, migration, and invasion in vitro.
Conclusions:
- Targeting CD44 presents a potential therapeutic strategy to prevent osteosarcoma metastasis.
- Inhibition of CD44 may improve clinical outcomes for osteosarcoma patients.
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