The LNP formulation loaded with eSpCas9 mRNA (m1Ψ) and TRAC sgRNA (at the molar ratio of 1:10) is prepared using generic LNP formulation with main ionizable lipid of ALC0315. The payload eSpCas9 mRNA (m1Ψ) has all the U bases 100% modified with N1-methyl-pseudoUridine. HPLC purified SafeEdit sgRNA with modifications targeting the first exon of the constant chain of the TCRα gene (TRAC), which enhances CAR-T cell potency and persistence by utilizing the endogenous transcriptional control of TCR gene. This formulation is a good control for testing if ALC0315 LNP is a good potential formulation for your CRISPR gene know out product under research in your in vitro and in vivo experiment model.