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The Genome Editing Core (GEC) provides DRC investigators with resources to manipulate the genome of human cell lines and laboratory mice with the aim of studying the genetics underlying diabetes and its complications. To facilitate these genetic studies, the GEC maintains a centralized facility for the generation and propagation of patient-derived induced Pluripotent Stem cells (iPSC). iPS cells hold tremendous potential for disease modeling applications as these cell lines retain the same genetic make-up as the patient's somatic cells targeted for reprogramming. The addition of gene editing tools (such as CRISPR/Cas 9) with iPSCs enables the study of genetic perturbations within a fixed genetic background, overcoming limitations inherent to studies based on genetically heterogeneous patient samples. The GEC also maintains a facility for the generation of transgenic mouse models to aid in studying the genetic mechanisms underlying diabetes in vivo. The mouse modeling facility uses CRISPR/Cas9 or lentiviral RNAi constructs to introduce genome modifications to a variety of mouse lines in order to establish new models of the disease within a living mammalian system.
Induced pluripotent stem cells (iPS cells), generated by transcription factor-dependent nuclear reprogramming of differentiated somatic cells, are pluripotent stem cell lines that can be propagated indefinitely in culture and maintain the potential to differentiate into any cell type in the body. As iPS cells retain the same genetic make-up as the somatic cell targeted for reprogramming, these cells hold tremendous promise for uncovering novel genetic and biochemical factors that underlie diseases with complex and poorly understood genetic influences, such as diabetes. The newly established iPS Core maintains a centralized facility for the reliable and consistent generation and propagation of reprogrammed iPS cells for use in cutting-edge research into the molecular and cellular pathologies underlying diabetes and its complications.
IPS Services Offered:
This Core enables investigators to manipulate the genome of laboratory mice in order to generate the most relevant experimental models to understand the genetics and mechanisms underlying diabetes and its complications. The GEC blends Joslin's research expertise in mouse models for diabetes and the generation of genetically modified mice.
The GEC will support the production of genetically modified mice, tailored to investigators' experimental questions by performing pronuclear and intracytoplasmic microinjections. If necessary, the Core can also provide lentiviral transgenesis services for generating mice that carry transgenes mediating inducible or constitutive silencing of target genes of interest by RNAi. The primary method of genetic modification is the CRISPR/Cas9 complex.
The Core will perform mouse genome editing using a variety of targeted edits made possible through CRISPR/Cas9. This includes creating knockout mice by generating a random mutation following a double stranded break (DSB) and non-homologous endpoint joining (NHEJ). For more specific modifications, the use of DNA templates can effect sequence insertion or replacement for knock-in models, for example. Other methods of influencing gene activation or repression are available and can be used upon request.
For questions about the GE Core and its iPSC services, please contact Amy Wagers: Amy.Wagers@joslin.harvard.edu or email: iPSCore@joslin.harvard.edu. For questions about CRISPR modified mice, please contact Taylor Roberts (Taylor.Roberts@joslin.harvard.edu) or Stephan Kissler (Stephan.Kissler@joslin.harvard.edu).
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