NHLBI iPSC Core Facility

National Heart, Lung, And Blood Institute

Investigators

Linked publications & trials

Abstract

In FY23, we received significant increase of requests for iPSC gene-editing services including 6 gene knockout/deletion, 18 gene-correction, 18 mutation or short-tag knock-in, 1 safe harbor knockin, and 4 reporter knockin-in projects. In total, we completed 47 gene-editing projects which are more than double of annual gene-editing projects we completed in the past 5 years. This trend reflects that NIH labs become more interested in using gene-editing technologies to create sophisticated iPSC disease models. In FY23, we also expanded our gene-editing services to PiggyBac-mediated transgenesis, which is an alternative and quick method to create reporter line and transgenic line. These iPSC gene-editing projects were done by Core Director, one staff scientist, and one biologist for NHLBI, NCATS, NEI, NHGRI, NINDS, and NIDDK labs. The genetically modified iPSC lines are being used as cell line models to study human diseases. In FY23, we continued to provide high-quality, integration-free iPSC generation services. When our users prefer or only have lymphoblastoid cell lines (LCL) that are available in many large cell repositories, we developed both Sendai Virus (SeV) and episomal vector mediated integration-free iPSC generation service. We now can use the same Cytotune 2.0 SeV kit and PBMC reprogramming protocol for LCL reprogramming. This simplifies the standard operating procedure (SOP) in our lab. In total, we reprogrammed 61 samples that included 10 skin fibroblast, 47 PBMC, and 4 LCL samples. These reprogramming projects were for NHLBI, NEI, NIMH, NHGRI, NCATS, NINDS, NIAID, and CC labs. The iPSC reprogramming projects were done by one biologist with the help from other Core staff. In FY23, we expanded our iPSC characterization services to include Scorecard assay, which takes less than 1 month to complete and can replace teratoma assay that usually takes more than 2 months. We continue to provide karyotyping, STR DNA fingerprinting, SeV qRT-PCR, mycoplasma test, and flow cytometry analysis of pluripotency markers. These services make a comprehensive package of standard iPSC characterization assays, which helped our users to publish their papers and share iPSC lines faster. Due to the popularity of our iPSC characterization services, >20% of our FY23 cost recovery was from these services. In FY23, we attracted NIA, NIAMS, NICHD, NIDDK, and NIDDS, in additional to NCATS, NIMH, NHGRI, and NEI to establish MOU and collect >$298,000 for NHLBI DIR to subsidize the personnel cost of iPSC Core. In addition, we collected >$267,000 service fees to completely cover the operating cost of iPSC Core and achieved double of budget recovery goal. Among the service fees we collected, 47% were from gene-editing services; 26% from iPSC generation services; 21% from iPSC characterization services. In FY23, we published 14 papers (https://www.ncbi.nlm.nih.gov/myncbi/jizhong.zou.1/bibliography/public/ ).