PUMA Gene and p53 Binding Site Knockout Cell Lines for Cancer Research

These cell lines are unique and useful tools for cancer research, specifically in the area of apoptosis. The most significant advantage of these cell lines is that they allow for the study of apoptosis mediated by PUMA, a protein that plays a key role in programmed cell death.

Description

The PUMA p53 binding site knockout cell lines (HCT-116 and DLD1) were created using AAV gene targeting technology to delete both copies of the PUMA p53 binding sites. These cells are isogenic compared to the parental cell lines and the deletion was confirmed through DNA sequencing. The PUMA knockout DLD1 cell line was also created using AAV gene targeting technology to delete both copies of the entire PUMA gene. These cells are also isogenic and are completely devoid of PUMA mRNA and protein expression. Homologous recombination is the only effective method for permanently deleting transcription factor binding sites.

Applications

- Cancer Research
- Drug Discovery

Advantages

- Useful and unique tools for studying cancer in the area of apoptosis mediated by PUMA.
- Isogenic compared to their parental lines
- The PUMA knockout DLD1 cell line is 100% devoid of PUMA mRNA and protein expression.

Invention Readiness

The technology is at the in vitro data stage of development. These knockout cell lines have been created using AAV gene targeting technology to delete either both copies of the PUMA p53 binding sites or both copies of the entire PUMA gene. The deletion of the p53 binding sites was confirmed by DNA sequencing, and the PUMA knockout cell lines were confirmed to be 100% devoid of PUMA mRNA and protein expression. Further studies would be needed to move beyond the in vitro data stage and validate these findings in more complex biological systems.

IP Status

Research Tool

Related Publication(s)

Wang, P., Yu, J., & Zhang, L. (2007). The nuclear function of p53 is required for PUMA-mediated apoptosis induced by DNA damage. Proceedings of the National Academy of Sciences, 104(10), 4054–4059. https://doi.org/10.1073/pnas.0700020104