Fabry Disease Model by CRISPR/Cas9 GLA KO

GLA gene knockout (KO) mice were used as Fabry disease model animals. These mice were generated in the Shanghai Model Organisms Center (Shanghai, China) using CRISPR/Cas9 technology. Genotyping was performed using DNA from collected tail samples to confirm the loss of the whole precursor-Gal gene encoding sequence (1400 bp). All mice used in the study were housed and bred in the Shanghai Model Organisms Center (Shanghai, China). During all experiments, mice had free access to clean food and water in a facility with a 12-h light/dark cycle. Animal experiments were authorized by the Animal Care and Use Committee of Shanghai Nanfang Model Biotechnology Co., Ltd (Shanghai, China) (2021-0012). Mice were randomly divided into three groups: a control group (PBS), a low-dose mRNA group (1 mg/kg), and a high-dose mRNA group (3 mg/kg). The mRNA encapsulated by LNP was injected into the mouse body via tail vein injection. The plasma, heart, liver, and kidneys of mice were collected at different times for testing α-Gal A activity and Lyso-Gb3 levels.
Male mice (wild-type) were housed in the Shanghai Model Organisms Center (Shanghai, China). The firefly luciferase mRNA (NCBI accession number M15077.1) encapsulated in LNP (1 mg/kg) was injected into the mouse body via tail vein injection. Fluorescence was observed at different locations and time points (0, 2, 6, 12, 24, 48, and 72 h) using a small animal imager (Clinx, IVScope 8000, Shanghai, China).