Publications

Samah W Awwad, Holly Barber, Josie Coulthard, Simon Lam, Alejandra Rojas, Nadia Gueorguieva, Michael Woods, Gabriel Balmus, Rimma Belotserkovskaya, Stephen P Jackson.

Nucleic Acid Research Volume 54, Issue 15, 27 August 2026.

We previously showed that loss of the RNA polymerase II Mediator subunits CDK8 and Cyclin C (CCNC) confers resistance to ATRi by suppressing transcription-dependent replication stress. To identify vulnerabilities that could be exploited to restore ATRi sensitivity in these resistant settings, we performed genome-wide CRISPR screens in wild-type and CDK8-deficient cells. These screens revealed impairment of epigenetic components HDAC3 or the PRC2 complex as synthetic vulnerabilities that enhance ATRi sensitivity, particularly in contexts of loss of CDK8 or CCNC.

Islam Md.S, Gautsch V.G., Belotserkovskaya R., Serrano-Benitez A., Buzafalvi D., Perisic O., Jackson S.P., Williams R.L.

BioRxiv

The Ser/Thr kinase ATM orchestrates cellular responses to DNA double-strand breaks (DSBs) and promotes DSB repair by homologous recombination. In this process, ATM is activated by DNA and the MRN (MRE11, RAD50, and NBS1) complex. Here we show that mutations of the conserved PIKK regulatory domain (PRD) within ATM’s kinase domain can confer a maximally active state that no longer requires MRN/DNA.

Bader A.S & Bushell M.

Nature Protocols Online ahead of print

DNA double-strand break (DSB)-induced mutagenesis is a critical contributor to a variety of human diseases; however, the ability to quantify these mutations is limited by the low sensitivity of current methodologies. Here we describe iMUT-seq, a technique that profiles DSB-induced mutations at high sensitivity and single-nucleotide resolution around endogenous DSBs, by coupling high-depth sequencing to restriction enzyme-inducible DSBs in human cell lines.

Zhao S, Palma-Chaundler CS, Engel CM, Cordes J, Nixdorf D, Luo MY, Kaya S, Suryo Rahmanto A, van den Heuvel D, Mackens-Kiani T, Weickert P, Lam S, Gupta V, Philippou-Massier J, Bagarić I, Bohlen J, Hewitt G, Luijsterburg MS, Beckmann R, Beli P, Nedialkova DD, Carnie CJ, Subklewe M, Jackson SP, Stingele J.

Molecular Cell 86(7), 1275-1292.e12.

Highlights

• CRISPR screens reveal cellular networks responding to azacytidine treatment

• The integrated stress response (ISR) drives azacytidine-induced cytotoxicity

• Incorporation of azacytidine into mRNA causes ribosome stalling and ISR activation

• RNF25 prevents ISR activation by ubiquitylating the small ribosomal subunit

Aird EJ, Serrano-Benitez A, Siegner SM, Cannavo E, Belotserkovskaya R, Gueorguieva N, Fielden J, Cullot G, Ammann S, Bader AS, Gupta V, Andrieux G, Raab R, Del Rey González M, Cathomen T, Cejka P, Corn JE, Jackson SP.

Nature Communications 17, Article number: 2743 (2026)

DNA double-strand breaks (DSBs) both pose threats to genome integrity and are commonly used for genome editing applications. Structural features of DSB ends play key roles in determining DNA repair pathway usage and outcomes during genome editing, but the cellular factors involved in these processes are only partially known. Through genome-wide CRISPRi screening, we identify ERCC6L2 as critical for repairing Cas12a-induced staggered DSBs but irrelevant for Cas9-induced blunt DSBs.

Aird EJ, Rabl J, Knuesel T, Groen K, Awwad SW, Korablev B, Scherpe L, Al-Herz W, Hupfer R, Recher M, Jackson SP, Hale BG, Corn JE.

Nature Cell Biology Online ahead of print.

Stimulation of the innate immune system by foreign RNA elicits a potent interferon response and can trigger cell death. The mechanisms by which cells balance a robust response with cell-intrinsic lethality are still being uncovered. Here, using genome-wide CRISPR–Cas9 genetic screens with triphosphorylated RNA stimulation, we discover that promyelocytic leukaemia (PML) nuclear body-localized speckled protein 110 (SP110) is a potent inhibitor of type 1 interferon-driven cell death.

Dogan H, Liptay M, Barbosa JS, Gogola E, Duarte AA, Schmid JA, Klebic I, Mutlu M, Siffert M, Francica P, Salguero I, van de Ven M, de Korte-Grimmerink R, Jackson SP, Jonkers J, Lopes M, Dibitetto D, Rottenberg S.

Oncogene 45, 491–504 (2026)

MDC1 is a key protein in DNA damage signaling. When DNA double-strand breaks (DSBs) occur, MDC1 localizes to the sites of DNA damage to promote the recruitment of other factors, including the 53BP1-mediated DSB repair pathway. By studying mechanisms of poly (ADP-ribose) polymerase inhibitor (PARPi) resistance in BRCA2; p53-deficient mouse mammary tumors, we identified a thus far unknown role of MDC1 in replication fork biology. Our results show that MDC1 localizes at active replication forks during normal DNA replication and regulates replication fork progression.

Hough SH, Jhujh SS, Awwad SW, Lewis OE, Lam S, Thomas JC, Mosler T, Bader A, Bartik L, McKee S, Amudhavalli S, Colin E, Damseh N, Clement E, Cacheiro P, Majumdar A, Smedley D, Fluss J, Giannini R, Thiffault I, Zagnoli Vieira G, Belotserkovskaya R, Smerdon SJ, Beli P, Galanty Y, Carnie CJ, Stewart GS, Jackson SP.

EMBO Molecular Medicine 18, 492–513, (2026)

Ubiquitin E3 ligases play crucial roles in the DNA damage response (DDR) by modulating the turnover, localization, activation, and interactions of DDR and DNA replication proteins. We performed a CRISPR-Cas9 knockout screen focused on ubiquitin E3 ligases and related proteins with the DNA topoisomerase I inhibitor camptothecin. This led us to establish that MAEA, a core subunit of the CTLH E3 ligase complex, is a critical regulator of homologous recombination and the replication stress response.