NEI Genetic Engineering Core

National Eye Institute

Investigators

Linked publications & trials

Abstract

During the past year, we have performed 135 procedures of electroporation and microinjection to produce multiple genetic engineered mouse strains for NEI and several other institutes on NIH campus including NIDCD, NICHD, NINDS, NIAMS and NIDDK through CRISPR-mediated mutagenesis. In addition, GEC produced 6 genetically modified human iPSC lines and reprogramed PBMC from two patients to iPSC using the Sendai virus system. Collaborating with a NEI team, we finished developing an allelic series of the mouse Nrl transcription factor aimed at defining alternative enhancers responsible for the expression of Nrl in the brain. A new reporter knockin mouse model of the Isg20 gene was created by our team in collaboration with a NEI lab, that is being used for the investigation of mechanisms of amyloid-beta (Aβ) build up in the plaques. Partnering with an immunology group in NIAMS, we designed a novel strategy generating TCR knockin mouse models and made progress in creating a knockin allele carrying cloned TRC alpha and beta chains that define a T-cell surface receptor responsible for Vitiligo, an autoimmune disease that causes the skin to lose its color, resulting in white or lighter patches. The similar strategy of generating knockin alleles of specific TCR clones can be used in the development therapeutic strategies in molecular immune therapy. Meanwhile, the engineered mutation in the mitochondria gene ND5 from last year’s work generated phenotype revealing its role in retinal degeneration. In the past year, GEC continue worked on human iPSC to reprogram human patient PBMCs to iPSC and to genetically modify the genome of human iPSC lines aimed at establishing human disease models in culture dish. In the area, we have successfully reprogramed additional PBMC samples from patients with USHER’s syndrome to iPSCs, providing valuable starting materials for the study of USHER’s syndrome. We also designed strategies and generated knockin and knockout alleles in human iPSC isolated from AMD patients for the study AMD etiology, and potential treatment. Genotyping operation: GEC genotyping services aided 10 laboratories in genomic DNA isolation of total ~10,000 samples, 2 laboratories in PCR genotyping of total 2,000 reactions and 13 laboratories in sample submission to Transnetyx. Cryopreservation and rederivation operations: GEC Cryopreservation services assisted a total of 15 laboratories of NEI in Sperm/Embryo cryopreservation, Re-derivation, colony expansion and strain rescue with a total of 60 projects. In addition, GEC performed 45 sperm cryopreservation projects for several labs from NIDCD and NINDS.