Publications

Reilly EL and Aitken SJ

Nature 642(8068):576-577

Instability of the genome is a hallmark of cancer that can arise when damaged DNA is carried across cell divisions, fuelling both small- and large-scale genetic variation in cells. Writing in Nature, Panagopoulos et al. present a multimodal platform that tracks single cells to uncover how DNA damage propagates unevenly through cell lineages. The authors’ approach reveals the origin and extent of differences in the traits and genomic stability of individual cells in each of the daughter and granddaughter generations produced from successive cell divisions.

Fielden J, Siegner SM, Gallagher DN, Schröder MS, Dello Stritto MR, Lam S, Kobel L, Schlapansky MF, Jackson SP, Cejka P, Jost M, Corn JE.

Nature 640(8060):1093-1102

The DNA damage response (DDR) is a multifaceted network of pathways that preserves genome stability. Unravelling the complementary interplay between these pathways remains a challenge. Here we used CRISPR interference (CRISPRi) screening to comprehensively map the genetic interactions required for survival during normal human cell homeostasis across all core DDR genes. We captured known interactions and discovered myriad new connections that are available online.

Awwad SW, Doyle C, Coulthard J, Bader AS, Gueorguieva N, Lam S, Gupta V, Belotserkovskaya R, Tran TA, Balasubramanian S, Jackson SP.

Nature Communications 16(1):480

ATR plays key roles in cellular responses to DNA damage and replication stress, a pervasive feature of cancer cells. ATR inhibitors (ATRi) are in clinical development for treating various cancers, including those with high replication stress, such as is elicited by ARID1A deficiency, but the cellular mechanisms that determine ATRi efficacy in such backgrounds are unclear. Here, we have conducted unbiased genome-scale CRISPR screens in ARID1A-deficient and proficient cells treated with ATRi.

Gang Goh C, Bader AS, Tran T-A, Belotserkovskaya R, D’Alessandro G and Jackson SP.

Nucleic Acids Research, gkae1163

HAP1 is a near-haploid human cell line commonly used for mutagenesis and genome editing studies due to its hemizygous nature. We noticed an unusual hypersensitivity of HAP1 to camptothecin, an antineoplastic drug that stabilizes topoisomerase I cleavage complexes (TOP1ccs). We have attributed this hypersensitivity to a deficiency of TDP1, a key phosphodiesterase involved in resolving abortive TOP1ccs.

Carnie CJ, Jackson SP and Stingele J.

Trends in Cell Biology 35(4), 316-329

DNA–protein crosslinks (DPCs) are highly toxic DNA lesions that are relevant to multiple human diseases. They are caused by various endogenous and environmental agents, and from the actions of enzymes such as topoisomerases. DPCs impede DNA polymerases, triggering replication-coupled DPC repair. Until recently the consequences of DPC blockade of RNA polymerases remained unclear. New methodologies for studying DPC repair have enabled the discovery of a transcription-coupled (TC) DPC repair pathway.

Lam S, Thomas JC and Jackson SP.

Genome Medicine 16:139 (2024)

Background CRISPR-Cas9 technology has revolutionised genetic screens and can inform on gene essentiality and chemo-genetic interactions. It is easily deployed and widely supported with many pooled CRISPR libraries available commercially. However, discrepancies between the reference genomes used in the design of those CRISPR libraries and the cell line under investigation can lead to loss of signal or introduction of bias. The problem is particularly acute when dealing with variant cell lines such as cancer cell lines.

Results Here, we present an algorithm, EXOme-guided Re-annotation of nuCleotIde SEquences (Exorcise), which uses sequence search to detect and correct mis-annotations in CRISPR libraries.

Lim JKM, Samiei A, Delaidelli A, de Santis JO, Brinkmann V, Carnie CJ, Radiloff D, Hruby L, Kahler A, Cran J, Leprivier G, Sorensen PH.

Cell Death & Disease 15(7):501.

Eukaryotic elongation factor 2 (eEF2) kinase (eEF2K) is a stress-responsive hub that inhibits the translation elongation factor eEF2, and consequently mRNA translation elongation, in response to hypoxia and nutrient deprivation. EEF2K is also involved in the response to DNA damage but its role in response to DNA crosslinks, as induced by cisplatin, is not known. Here we found that eEF2K is critical to mediate the cellular response to cisplatin.

Carnie CJ, Götz MJ, Palma-Chaundler CS, Weickert P, Wanders A, Serrano-Benitez A, Li HY, Gupta V, Awwad SW, Blum CJ, Sczaniecka-Clift M, Cordes J, Zagnoli-Vieira G, D'Alessandro G, Richards SL, Gueorguieva N, Lam S, Beli P, Stingele J, Jackson SP.

EMBO Journal 43, 2397 - 2423

The nucleoside analogue decitabine (or 5-aza-dC) is used to treat several haematological cancers. Upon its triphosphorylation and incorporation into DNA, 5-aza-dC induces covalent DNA methyltransferase 1 DNA-protein crosslinks (DNMT1-DPCs), leading to DNA hypomethylation. However, 5-aza-dC's clinical outcomes vary, and relapse is common. Using genome-scale CRISPR/Cas9 screens, we map factors determining 5-aza-dC sensitivity.