Oncotarget: CRISPR Used in Triple Negative Breast Cancer Research

In this week's cover paper of Oncotarget (Volume 12, Issue 12), entitled, "Frame-shift mediated reduction of gain-of-function p53 R273H and deletion of the R273H C-terminus in breast cancer cells result in replication-stress sensitivity," researchers used the CRISPR-Cas9 tool to analyze a p53 mutation in triple-negative breast cancer. The researchers from the City University of New York, Columbia University, and Weill Cornell Medical College wrote that both the C-terminal domain (CTD) and oligomerization domain (OD) of mtp53 R273H proteins are intact, and it is not clear if these regions are responsible for chromatin-based DNA replication activities. To examine the ability of mtp53 R273H to influence cell proliferation, DNA replication, and cell cycle progression of breast cancer cells, the researchers used the CRISPR-Cas9 tool to edit the C-terminal regions of the mtp53 gene. “CRISPR-Cas9 sgRNA editing of the C-terminal regions of the endogenous mtp53 gene were carried out so as to delete gene sequences that correspond to the OD and CTD regions.” The team generated breast cancer cells and edited CTD and OD regions of mtp53 R273H using the CRISPR-Cas9 tool. They then treated the cell populations with thymidine—to block cells at G1/S phase in the cell cycle. The researchers then compared the status and proliferation of the variants with the original cell line and observed changes in total cell lysates by western blot analysis. “We examined how changes in the level of mtp53 R273H level and/or deletion of the CTD, or OD plus CTD, region influenced cell proliferation, cell cycle progression, and chromatin association of mtp53, RPA, PCNA and MCM2.” Sign up for free Altmetric alerts about this article - https://oncotarget.altmetric.com/details/email_updates?id=10.18632%2Foncotarget.27975 DOI - https://doi.org/10.18632/oncotarget.27975 Full text - https://www.oncotarget.com/article/27975/text/ Correspondence to - Jill Bargonetti - bargonetti@genectr.hunter.cuny.edu Keywords - mutant p53, gain-of-function, oligomerization, DNA replication, frame-shift, breast cancer About Oncotarget Oncotarget is a bi-weekly, peer-reviewed, open access biomedical journal covering research on all aspects of oncology. To learn more about Oncotarget, please visit https://www.oncotarget.com or connect with: SoundCloud - https://soundcloud.com/oncotarget Facebook - https://www.facebook.com/Oncotarget/ Twitter - https://twitter.com/oncotarget YouTube - https://www.youtube.com/c/OncotargetYouTube/ LinkedIn - https://www.linkedin.com/company/oncotarget Pinterest - https://www.pinterest.com/oncotarget/ Reddit - https://www.reddit.com/user/Oncotarget/ Oncotarget is published by Impact Journals, LLC please visit https://www.ImpactJournals.com or connect with @ImpactJrnls Media Contact MEDIA@IMPACTJOURNALS.COM 18009220957