Screening for Key Genes of T Cell Uptake of CAMP LNP via CRISPR Screening

CAMP (Cardiolipin-mimic lipid) lipid nanoparticle (LNP) can transfect T cell selectively without antibody modification. The multiple faceted surface, thinner lamellar and higher rigidity facilitate the selective transfection. However, the molecular mechanism underlying this selective property remains unclear. We performed membrane protein CRISPR KO screen in the mouse T cell line CT237. Subsequently, we transfected the cells with PL40 LNP (a representative CAMP LNP) encapsulating mCherry mRNA. We then divided the cells into four equal parts based on the intensity of mCherry expression. We compared the two parts with the lowest and highest expression, and then screened for membrane proteins specifically expressed in T cells. Finally, we identified that CD44, CD52 and IL18rap might be the key genes associated with the uptake of PL40-LNP.