NEI Genetic Engineering Core

National Eye Institute

Investigators

Linked publications, trials & patents

Abstract

Summary Coming out of the pandemic of Covid-19, we accomplished total of 53 genetic engineering projects either through CRISPR-mediated mutagenesis in mouse or classic homologous recombination in mouse embryonic stem cells, for the studies of physiology or pathology of target genes. It is often desirable to have mutations of human genetic conditions replicated in mouse so that the diseases can be modeled. In the last year, several such models are being worked on at GEC in collaboration with investigators at NEI, NIDCD and NICHD. One of the NADH dehydrogenase coding variants was looked at for its relationship with retinal degeneration. We in collaboration with NEI scientists performed base editing in mouse to explore the feasibility of establishing a mouse model for the disease. We established a conditional allele for one of the coloboma genes, the ZFP503, for the modeling of genetically arising coloboma conditions in human pediatric patients. We also created a missense mutation of the Cp110 gene to study the pathologic role of this disease allele. RPGR gene mutations in human account for about 70 percent of all cases of X-linked retinitis pigmentosa (RP). The rd9 allele of RPGR is caused by a 32bp insertion in the highly repetitive ORF15 domain of the RPGR gene. We in collaboration with NEI investigators performed germline gene therapy by indel mutagenesis with CRISPR/Cas9 and successfully restored normal open reading frame of the rd9. Pups of the F0 founders with edited RPGR gene developed normal photoreceptors and vision. Genotyping operation: GEC genotyping services aided total of 18 laboratories most of which are from NEI intramural program. * isolated DNA from 11,296 mouse tail biopsy samples, almost doubled from last years number * Performed 5,419 PCR reactions to genotype mice in the facility, also doubled last years work load. * acted on behalf of several laboratories sending mouse tissue samples to Transnetyx for genotyping services. Cryopreservation and rederivation operations: GEC Cryopreservation services assisted many laboratories from NEI in cryopreserving 30 lines, rederivation of 5 line, and rescuing of 3 lines.