Gene editing in induced pluripotent stem cells (iPS cells) has within the last couple of years emerged as a valuable tool. One of the most powerful applications is the generation of isogenic lines for patient specific disease modelling. Such isogenic iPS cell lines have turned out to be a sensitive tool that have the potential to uncover even subtle phenotypical changes caused by single mutations associated with a certain disease.
Alzheimer’s disease (AD) is the most common form of dementia in the elderly. Except for a few known familiarly disease-causing mutations, 99% of all cases are categorized as sporadic late-onset with only poorly understood pathomechanisms. However, several risk genes have been identified, such as the ApoE gene.
EBiSC is proud to announce the first human complete set of isogenic lines with the main ApoE genotypes; ApoE 2/2 (BIONi010-C-6), ApoE 3/3 (BIONi010-C-2) and ApoE 4/4 (BIONi010-C-1) generated by Bioneer A/S. Genotypes were introduced into a control iPS cell line from a 19 years old male individual (ApoE3/E4, BIONi010-C) using CRISPR-Cas9 technology. Moreover, Bioneer A/S has also generated an isogenic ApoE knockout line (BIONi010-C-3). Previous publication from the group of Nobel laureate Thomas Suedhof (Huang et al., 2017) showed the huge potential of different ApoE genotypes for AD modeling. By applying recombinant ApoE expression (ApoE2, ApoE3 or ApoE4) to human neurons differentiated by enforced overexpression of the ngn2 gene, Huang et al. were able to elucidate a possible ApoE related pathomechanism for AD in vitro. Due to the extensive request for the set of ApoE lines that is already available at EBiSC, similar sets of isogenic ApoE lines are currently being generated from three other iPS cell lines: One derived from a healthy female individual (ApoE3/E3 genotype) as well as one from both a female and a male AD patient diagnosed with an ApoE 4/4 genotype. These lines will also be available through EBiSC later this year.
Other generated iPS cell lines of interest to researchers in the AD field include an iPS cell line, where Bioneer targeted the TREM2 gene and inserted either a homozygous R47H SNP (BIONi010-C-7) or a homozygous T66M SNP (BIONi010-C-8) - both risk SNPs associated with AD. Additionally, iPS cell lines targeting the CD33 locus have been generated: a knockout and a variant lacking exon 2 (BIONi010-C-9 and BIONi010-C-5 respectively).
EBiSC is developing tools to accelerate neuronal differentiation of iPSCs
Bioneer is currently working on combining their initial isogenic sets of ApoE lines with doxycycline controllable expression of Ngn2. The aim is to develop a robust platform for fast and effective neuronal disease models that can significantly shorten, improve and lower the costs of the differentiation process of iPSC lines.
“We believe that these combined lines, where differentiation into neurons can be done within weeks instead of months, will be very valuable tools in our efforts of generating high qualitative disease models,” says Bioneer’s Director of Research and Development, Dr. Christian Clausen.
Please confirm your country of origin from the list below.