In partnership with the iPSC Neurodegenerative Disease Initiative, a project from the Center for Alzheimer’s and Related Dementias (NIH), and with support from the Chan Zuckerberg Initiative, The Jackson Laboratory now offers a catalog of human induced pluripotent stem cells (iPSCs) for Alzheimer’s and Related Dementias (ADRDs), ALS, and other neurodegenerative disorders.
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We have selected more than 100 ADRD-associated variants across 73 genes through automated identification and input from leading geneticists. CRISPR-Cas technology was employed to generate heterozygous and homozygous genotypes of these single nucleotide variants (SNVs) in the KOLF2.1J cell line†, along with mutation-corrected isogenic “revertant” controls for each mutation. When paired together, these sets offer a broad, quality-controlled toolbox of ADRD cell lines along with genetically-matched controls for highly efficient screening and reproducible results. For more details on the selection of variants, click HERE.
JAX groups cell lines into Distribution Sets. The core cell lines in a Distribution Set are composed of the heterozygous mutation, the homozygous mutation and the revertant line. The parental cell line, KOLF2.1J, should be used as a reference cell line for all Distributions Sets. Individual cell lines can be purchased but we recommend using the complete set of cell lines along with the parental cell line for well-controlled studies.
The parental cell line used for the generation of this catalog’s gene-edited variants is the KOLF2.1J cell line. This cell line was selected based on a variety of characteristics including its genomic sequence, genome stability in culture, cellular characteristics (i.e., pluripotency) and amenability to gene editing protocols. The KOLF2.1J cell line was derived from the HPSI0114i-kolf_2 line through extensive molecular and cellular characterization efforts, including the correction of a pathogenic mutation in ARID2.
Our panel of SNVs was selected based on coding changes from inherited ADRD identified in the literature based on linkage and segregation of variants in families. In total, 134 variants spread across 73 genes were chosen to be engineered into the KOLF2.1J parental cell line. Each CRISPR-edited heterozygous and homozygous SNV cell line has been characterized through targeted sequence analysis (PCR-Sanger) and quality controlled using karyotype, genomic wide copy number variant and molecular pathogen analyses. We offer both heterozygous and homozygous cell lines to enable assessments of allelic dosage effects.
Non-Desired Mutations: CRISPR-based editing procedures can inadvertently produce mutations outside the sequence region used in the characterization process. These are frequently referred to as “off-target” effects. These mutations are typically distant from the region being targeted, most often residing on other chromosomes. In contrast, the same process can yield mutations in the region of interest that can go undetected by the targeted PCR-Sanger sequencing process. Even current “next-generation” sequencing technologies and their associated computational algorithms can fail to identify these aberrant “on-target” mutations. Most “off-target” effects have very similar sequences when compared to the desired targeted sequence and can be mitigated in the design strategy. In contrast, the “on-target” aberrant mutations are less predictable. These on- and off-target mutations can lead researchers to inappropriately assign functional significance to the engineered SNV, even when parental controls are used.
Rescue Mutation: “Hidden” undesired mutations can lead a user of an engineered cell line to misattribute phenotype causation to the declared SNV when the true cause is the uncharacterized mutation elsewhere in the engineered cell line genome. To reduce the risk that a researcher is misled by one or more of the above non-desired mutations in the SNV-carrying cell line, we have created precise, efficient controls. The SNV mutation-carrying cell line has undergone another round of gene engineering to revert the gene back to a 'wild type' status. We refer to these cell lines as “revertants” and they represent a high-value control for your in vitro experiments. If one detects a phenotype in the SNV-containing cell line compared to the parental control but the “revertant” continues to produce the same phenotype as the SNV cell line, the phenotype is likely caused by one, or several, undesired mutations.
We continue working to provide you the best tools available. We will be adding additional cell lines to our existing Distribution Sets, including whole gene knock-outs, SNV-containing and wild type genes fused to the multifunction HaloTag sequence. These accessory alleles will augment the utility of the “Trio” Distribution Sets.
To ensure cell survival and proliferation, JAX iPSCs should be thawed using the standard protocol provided below. Once cells have recovered and grow in log phase, they can be switched to alternative protocols. We suggest freezing some vials before changing any culture conditions.
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Our catalog of iPSCs is now available to for-profit and commercial customers. Please review the MTA above for applicable terms and conditions.
†The Kolf2_C1 cell line is owned by Genome Research Limited.
Licensing Information
+1.207.288.6470 (US)
+1.207.288.5845 (International)
TechTran@jax.org
This innovative catalog has been made possible with funding from the Chan Zuckerberg Initiative and our cell development partner the National Institute of Health.
