The LUMC human iPSC Hotel: Generation of research grade human iPSCs and beyond

Gene editing

Homology directed repair of small-scale mutations

Proper validation of hiPSC-based disease models requires comparison of the phenotypes between patient-derived disease lines and isogenic control lines, i.e. cells with the same genetic background in which the causal mutation was repaired. 

Our service

Cas9-mediated repair of mutations in your hiPSC line of choice (2 independent clones per disease line*).  It includes design and testing of three different sgRNAs, repair of the genomic mutation using ssODNs and Cas9/sgRNA ribonucleoproteins (RNPs), and pluripotency analysis of two selected isogenic sub-clones per disease clone. Off-target analysis and karyotyping is not included in this service. 

Info and requirements

This service is only available for hiPSC lines (passage >10 and <20) cultured in mTESR-E8 or Essential 8 that are mycoplasma-negative, and karyotypically normal (as assessed by G-banding or comparable methods). The hiPSC hotel has the right to decline the request on basis of the DNA sequence of the mutated genomic region. Requests should contain all required information and will be handled on a first-come-first-serve basis. Capacity is limited. 

The service is free of charge for LUMC researchers. External non-profit as well as commercial parties should first send an inquiry. For additional information or inquiries please contact c.m.a.h.freund@lumc.nl or h.mikkers@lumc.nl

*a second disease clone will be edited only if repair of the first clone was successful