Gene targeting in mouse has dramatically improved our understanding of the functions of genes in vivo in health and diseases. However, it is a time-consuming, laborious, and expensive process. The recent groundbreaking development of genome editing technologies has enabled direct manipulation of the genome in mouse zygotes (in vivo genome editing), thereby providing new avenues for simple, convenient, highly efficient and ultra-rapid production of knockout or knockin mice without ES cells. We focus on functional significance of multiple human rare variants in glutamate transporter EAAT1 found in glaucoma patients. To achieve this in vivo, we have generated several knockin mouse lines carrying precisely modified these rare variants with highly active Platinum TALENs and oligo DNA donors. We have also generated knockin mice carrying a 6.5kb gene cassette containing EGFP reporter with CRISPR/Cas and targeting vector for fine tracing of specific-cell lineage. Because the efficiency of our method is quite high, several germline-competent knockin founders can be obtained within a month by single microinjection. Taken together, in vivo genome editing technology revolutionizes mouse gene targeting and provides exciting opportunities for the functional genomic and neurodevelopmental researches in vivo.