A high-density CRISPR activation platform for mapping cancer dependencies and resistance pathways ex vivo and in vivo

Diepstraten, ST; Deng, Y; Potts, MA; Heidersbach, A; K&#246;nig, C; Dorighi, KM; Tai, L; Kueh, AJ; Whelan, L; Chang, C; Brown, F; Kelly, GL; Fortin, JP; Haley, B; La Marca, JE; Herold, MJ

Author(s): Diepstraten, ST; Deng, Y; Potts, MA; Heidersbach, A; König, C; Dorighi, KM; Tai, L; Kueh, AJ; Whelan, L; Chang, C; Brown, F; Kelly, GL; Fortin, JP; Haley, B; La Marca, JE; Herold, MJ;
Details: Publication Year 2026-09-18,Volume 12,Issue #38,Page eaec0722
Journal Title: Science Advances
Abstract: CRISPR activation (CRISPRa) enables precise up-regulation of gene expression for ex vivo and in vivo applications. However, a lack of scalable, high-coverage tools has limited comprehensive genetic screening in murine models. Here, we introduce Partita, a next-generation mouse whole-genome CRISPRa sgRNA platform, designed for unparalleled efficiency in gene activation studies. Partita uses a high-density targeting strategy, deploying 10 sgRNAs per transcriptional start site, structured into five gene class-specific sublibraries to maximize transcriptional induction. To demonstrate the capabilities of Partita, we performed a series of large-scale screens: an in vitro enrichment/depletion screen, whole-genome CRISPRa screens in a double-hit lymphoma model to uncover resistance factors to proapoptotic drugs (venetoclax, nutlin-3a, and etoposide) and an in vivo screen to identify accelerators of MYC-driven lymphomagenesis. Each experiment revealed both expected and unexpected regulators, with high validation rates. By enabling robust gain-of-function screening, Partita unlocks new avenues for functional genomics and expands the toolkit for discovering key drivers of biological processes across diverse research fields.
Publisher: AAAS
Keywords: Animals; Mice; *Drug Resistance, Neoplasm/genetics; *CRISPR-Cas Systems; *Clustered Regularly Interspaced Short Palindromic Repeats; Gene Expression Regulation, Neoplastic; Humans; Lymphoma/genetics/drug therapy; Proto-Oncogene Proteins c-myc/genetics; RNA, Guide, CRISPR-Cas Systems/genetics; *Neoplasms/genetics
Research Division(s): Blood Cells and Blood Cancer; Infection and Global Health; New Medicines and Diagnostics
PubMed ID: 42758820
Publisher's Version: https://doi.org/10.1126/sciadv.aec0722
Open Access at Publisher's Site: https://doi.org/10.1126/sciadv.aec0722
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2026-10-02 08:18:56

Last Modified: 2026-10-02 08:19:08