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Life sciences · Journal article
Sabaragamuwa University Journal · October 15, 2026
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CD36 (Cluster of Differentiation 36) is a transmembrane glycoprotein that plays a crucial role in various cellular processes, and has been implicated in cancer progression, metastasis, chemoresistance, and radio-resistance. CD36 has recently gained attention as a potential therapeutic target in cancer treatment. CRISPR/Cas9 technology has revolutionized genome editing, enabling precise modifications of specific genes. This study aims to design a CRISPR-based toolkit for the effective knockout of the CD36 gene in human cancer cells. A total of 66 sgRNAs were designed using ATUM (Gene-GPSTM), Edit-R CRISPR Design Tool (Horizon), and CHOPCHOP (v.3), and three sgRNAs were selected based on the target location, on-target efficiency, and minimized off-target effects. The designed sgRNAs were synthesized and cloned into the pSpCas9(BB)-2A-Puro vector, and the recombinant plasmids were verified through colony PCR and sequencing. Donor templates were designed to incorporate an EcoRI restriction site and a 6xHis-tag at the double-stranded break. The sgRNA-cloned recombinant vectors, along with their corresponding donor templates, will be co-transfected into cancer cells in vitro to facilitate the knockout of CD36. This CRISPR/Cas9 knockout strategy provides a robust approach to elucidate the role of CD36 in cancer development and holds potential as a targeted therapy for CD36 in cancer.