NEI Genetic Engineering Core

National Eye Institute

Investigators

Linked publications, trials & patents

Abstract

Summary During the past year, we have worked on 60 genetic engineering projects through CRISPR-mediated mutagenesis in mouse, which were initiated by investigators of NEI, NIDCD, NICHD, NINDS, NIAMS, NIGMS, NIDDK and NCI as well as Mayo Clinic. Collaborating with NINDS and Mayo Clinic researchers, we established a CRISPRi mouse line with an enhanced dCas9-KRAB configuration. This novel CRISPRi allele effectively repressed target gene expression in ES cells, 3D eye cup cultures, and mouse brains. Our mouse model knockout of the Spin4 gene mirrored human overgrowth syndromes as confirmed by NICHD's study. Partnering with an NEI group, we genetically modified the ND5 gene in female mitochondria, revealing its role in retinal degeneration. Additionally, we generated two mouse models of Leber Congenital Amaurosis-like conditions with missense mutants in the RPE65 gene. Furthermore, we created an allelic series of PNPLA6 gene mutations, showcasing their pathological implications in retinopathy. Lastly, in collaboration with another NEI team, we successfully developed a mouse model for Bietti Crystalline Dystrophy. Genotyping operation: GEC genotyping services aided 11 laboratories in genomic DNA isolation of total 12,607 samples, 5 laboratories in PCR genotyping of total 4,162 reactions and 8 laboratories in sample submission to Transnetyx. Cryopreservation and rederivation operations: GEC Cryopreservation services assisted a total of 10 laboratories of NEI in Sperm/Embryo cryopreservation, Re-derivation, colony expansion and strain rescue with a total of 53 projects. In addition, we also performed 11 cryopreservation projects for 3 labs from NIDCD and NINDS.