Interstrand crosslinks (ICLs) are highly toxic DNA lesions that prevent transcription and replication by inhibiting DNA strand separation. ICLs were among the first chemotherapeutic agents used, and their use has evolved over the years. Recent studies have provided insights into how cells repair ICLs, particularly through the Fanconi anemia (FA) pathway, which involves nucleases, helicases, and other DNA-processing enzymes. FA is a rare genetic disorder characterized by sub-fertility, congenital abnormalities, bone marrow failure, and an elevated risk of hematological and squamous cell cancers. The FA pathway coordinates multiple repair systems, including homologous recombination (HR), nucleotide excision repair (NER), and translesion synthesis (TLS). The FA core complex, composed of FANCA, FANCB, FANCC, FANCE, FANCF, FANCG, and FANCL, is central to ICL repair. FANCM, a structure-specific DNA-binding protein, recognizes ICLs and recruits the FA core complex to chromatin. Monoubiquitylation of FANCD2 and FANCI by the FA core complex facilitates their binding to damaged sites. Nucleases like FAN1, MUS81–EME1, XPF–ERCC1, and SLX1–SLX4 play crucial roles in ICL repair by cleaving the DNA backbone adjacent to the crosslink. HR is important for ICL repair, especially at stalled replication forks, where it promotes fork regression and the formation of chicken foot structures. Non-homologous end joining (NHEJ) is not essential for ICL repair but can be suppressed by the FA pathway to prevent spurious ligation between non-homologous chromosomes. Understanding ICL repair mechanisms has implications for improving the clinical use of ICL-inducing chemotherapeutic agents, such as carboplatin and platinum compounds, by modulating cell sensitivity and reducing toxicity. Strategies to enhance ICL repair in normal tissue and inhibit repair in tumors could improve therapeutic outcomes.Interstrand crosslinks (ICLs) are highly toxic DNA lesions that prevent transcription and replication by inhibiting DNA strand separation. ICLs were among the first chemotherapeutic agents used, and their use has evolved over the years. Recent studies have provided insights into how cells repair ICLs, particularly through the Fanconi anemia (FA) pathway, which involves nucleases, helicases, and other DNA-processing enzymes. FA is a rare genetic disorder characterized by sub-fertility, congenital abnormalities, bone marrow failure, and an elevated risk of hematological and squamous cell cancers. The FA pathway coordinates multiple repair systems, including homologous recombination (HR), nucleotide excision repair (NER), and translesion synthesis (TLS). The FA core complex, composed of FANCA, FANCB, FANCC, FANCE, FANCF, FANCG, and FANCL, is central to ICL repair. FANCM, a structure-specific DNA-binding protein, recognizes ICLs and recruits the FA core complex to chromatin. Monoubiquitylation of FANCD2 and FANCI by the FA core complex facilitates their binding to damaged sites. Nucleases like FAN1, MUS81–EME1, XPF–ERCC1, and SLX1–SLX4 play crucial roles in ICL repair by cleaving the DNA backbone adjacent to the crosslink. HR is important for ICL repair, especially at stalled replication forks, where it promotes fork regression and the formation of chicken foot structures. Non-homologous end joining (NHEJ) is not essential for ICL repair but can be suppressed by the FA pathway to prevent spurious ligation between non-homologous chromosomes. Understanding ICL repair mechanisms has implications for improving the clinical use of ICL-inducing chemotherapeutic agents, such as carboplatin and platinum compounds, by modulating cell sensitivity and reducing toxicity. Strategies to enhance ICL repair in normal tissue and inhibit repair in tumors could improve therapeutic outcomes.