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Abstract Cysteine-rich receptor-like-kinases (CRKs), a transmembrane subfamily of receptor-like kinase, play crucial roles in plant adaptation. As such cotton is the major source of fiber for the textile industry, but environmental stresses are limiting its growth and production. Here, we have performed a deep computational analysis ofCRKsin fiveGossypiumspecies, includingG. arboreum(60 genes), G. raimondii(74 genes), G. herbaceum(65 genes), G. hirsutum(118 genes), andG. barbadense(120 genes). All identified CRKs were classified into 11 major classes and 43 subclasses with the finding of several novel CRK-associated domains includingALMT, FUSC_2, Cript, FYVE,andPkinase. Of these,DUF26_DUF26_Pkinase_Tyrwas common and had elevated expression under different biotic and abiotic stresses. Moreover, the 35 land plants comparison identified several newCRKsdomain-architectures. Likewise, several SNPs and InDels were observed in CLCuD resistantG. hirsutum. The miRNA target side prediction and their expression profiling in different tissues predictedmiR172as a major CRK regulating miR. The expression profiling ofCRKsidentified multiple clusters with co-expression under certain stress conditions. The expression analysis under CLCuD highlighted the role ofGhCRK057, GhCRK059, GhCRK058, and GhCRK081in resistant accession. Overall, these results provided primary data for future potential functional analysis as well as a reference study for other agronomically important crops.