{"doi":"10.1101/2020.03.30.015271","title":"A multiplexed bioluminescent reporter for sensitive and non-invasive tracking of DNA double strand break repair dynamics <i>in vitro</i> and <i>in vivo</i>","abstract":"ABSTRACT Tracking DNA double strand break (DSB) repair is paramount for the understanding and therapeutic development of various diseases including cancers. Herein, we describe a multiplexed bioluminescent repair reporter (BLRR) for non-invasive monitoring of DSB repair pathways in living cells and animals. The BLRR approach employs secreted Gaussia and Vargula luciferases to simultaneously detect homology-directed repair (HDR) and non-homologous end joining (NHEJ), respectively. BLRR data are consistent with next-generation sequencing results for reporting HDR (R 2 = 0.9722) and NHEJ (R 2 = 0.919) events. Moreover, BLRR analysis allows longitudinal tracking of HDR and NHEJ activities in cells, and enables detection of DSB repairs in xenografted tumours in vivo. Using the BLRR system, we observed a significant difference in the efficiency of CRISPR/Cas9-mediated editing with guide RNAs only 1-10 bp apart. Moreover, BLRR analysis detected altered dynamics for DSB repair induced by small-molecule modulators. Finally, we discovered HDR-suppressing functions of anticancer cardiac glycosides in human glioblastomas and glioma cancer stem-like cells via inhibition of DNA repair protein RAD51 homolog 1 (RAD51). The BLRR method provides a highly sensitive platform to simultaneously and longitudinally track HDR and NHEJ dynamics that is sufficiently versatile for elucidating the physiology and therapeutic development of DSB repair.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2020,"id":129261,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9534,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":392901,"name":"Elie Tabet","orcid":"0000-0002-6905-1047","position":1,"is_corresponding":false},{"id":51295,"name":"Kelsey Pinkham","orcid":null,"position":2,"is_corresponding":false},{"id":511688,"name":"Cintia Carla da Hora","orcid":null,"position":3,"is_corresponding":false},{"id":511689,"name":"Jason Cheng-Yu Chang","orcid":null,"position":4,"is_corresponding":false},{"id":60476,"name":"Steven Lin","orcid":"0000-0002-7542-4663","position":5,"is_corresponding":false},{"id":489930,"name":"Christian E. Badr","orcid":"0000-0002-7212-5975","position":6,"is_corresponding":false},{"id":266751,"name":"Charles Pin‐Kuang Lai","orcid":"0000-0001-8050-7060","position":7,"is_corresponding":false},{"id":511687,"name":"Jasper Che-Yung Chien","orcid":null,"position":0,"is_corresponding":true}],"reference_count":74,"raw_metadata":null,"created_at":"2026-07-18T23:15:45.920615Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}