PXD064062

PXD064062 is an original dataset announced via ProteomeXchange.

Title	Stable Cas9 Expression on Cell Growth Regulation Through mTORC2 Activation Facilitation
Description	CRISPR, widely used for gene editing, relies on bacterial endonucleases like Cas9 to study gene functions and develop therapies. However, its potential effects on mammalian cellular responses and behavior remain unclear. Here, we systematically profiled effects of stable Cas9 expression on growth of 30 cell lines spanning 10 cancer types, observing growth alterations in 12 of them. To uncover the underlying mechanisms, we established the SpCas9 interactome in DU145 and MDA-MB-231 cells, both of which exhibited Cas9-increased growth phenotypes, and identified ribosomal proteins as the top shared interactors. Mechanistic studies revealed that ribosomal proteins such as RPL26 and RPL23a bind Sin1, a key mTORC2 component, relieving Sin1-PH-mediated suppression of the mTOR kinase domain leading to mTORC2 activation. Notably, SpCas9 interacts with both RPL26/RPL23a and Sin1, acting as a "glue" to stabilize their association and enhances mTORC2 activation, even in the absence of growth factors. Our study systematically characterizes Cas9’s effects on cell growth regulation and uncovers a novel Cas9-ribosome-mTORC2 signaling axis that promotes cell growth. These findings underscore the need to consider unintended cellular effects in CRISPR applications and highlight the importance of engineering safer Cas9 variants for biomedical research and clinical therapies.
HostingRepository	PRIDE
AnnounceDate	2025-10-06
AnnouncementXML	Submission_2025-10-05_16:41:38.086.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Emily Wilkerson
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Eclipse; Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2025-05-19 09:44:33	ID requested
⏵ 1	2025-10-05 16:41:38	announced

Yu L, Jin Y, Chen J, Zhu Z, Su S, Wilkerson EM, Gongora J, Cloer EW, Major MB, Liu P, Stable Cas9 expression regulates cell growth by facilitating mTORC2 activation. Nucleic Acids Res, 53(18):(2025) [pubmed]

10.1093/nar/gkaf965;

submitter keyword: DU145, MDA-MB-231

Pengda Liu
contact affiliation	Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA Department of Biochemistry and Biophysics, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA
contact email	pengda_liu@med.unc.edu
lab head
Emily Wilkerson
contact affiliation	Washington University School of Medicine
contact email	emilyz@wustl.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/10/PXD064062

PRIDE project URI

• PRIDE
  1. PXD064062
     1. Label: PRIDE project
     2. Name: Stable Cas9 Expression on Cell Growth Regulation Through mTORC2 Activation Facilitation