A new genetic tool to improve immune-compromised mouse models: Derivation and CRISPR/Cas9-mediated targeting of NRG embryonic stem cell lines
| Autor: | Teresa D'Altri, Aurélie Baudet, Cord Brakebusch, Javier Martin Gonzalez, Sahar Abelechian, Kasper Bonderup, Jörg Cammenga, Gunnar Juliusson, Bo T. Porse |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Fertilization in Vitro Biology Models Biological Germline Cell Line Immunocompromised Host Mice 03 medical and health sciences Endocrinology Genome editing Mice Inbred NOD Genetics Animals CRISPR Induced pluripotent stem cell Embryonic Stem Cells Gene targeting Cell Biology Embryonic stem cell Cell biology GATA2 Transcription Factor Haematopoiesis 030104 developmental biology Gene Targeting CRISPR-Cas Systems Stem cell |
| Zdroj: | genesis. 56:e23238 |
| ISSN: | 1526-954X |
| DOI: | 10.1002/dvg.23238 |
| Popis: | Development of human hematopoietic stem cells and differentiation of embryonic stem (ES) cells/induced pluripotent stem (iPS) cells to hematopoietic stem cells are poorly understood. NOD (Non-obese diabetic)-derived mouse strains, such as NSG (NOD-Scid-il2Rg) or NRG (NOD-Rag1-il2Rg), are the best available models for studying the function of fetal and adult human hematopoietic cells as well as ES/iPS cell-derived hematopoietic stem cells. Unfortunately, engraftment of human hematopoietic stem cells is very variable in these models. Introduction of additional permissive mutations into these complex genetic backgrounds of the NRG/NSG mice by natural breeding is a very demanding task in terms of time and resources. Specifically, since the genetic elements defining the NSG/NRG phenotypes have not yet been fully characterized, intense backcrossing is required to ensure transmission of the full phenotype. Here we describe the derivation of embryonic stem cell (ESC) lines from NRG pre-implantation embryos generated by in vitro fertilization followed by the CRISPR/CAS9 targeting of the Gata-2 locus. After injection into morula stage embryos, cells from three tested lines gave rise to chimeric adult mice showing high contribution of the ESCs (70%-100%), assessed by coat color. Moreover, these lines have been successfully targeted using Cas9/CRISPR technology, and the mutant cells have been shown to remain germ line competent. Therefore, these new NRG ESC lines combined with genome editing nucleases bring a powerful genetic tool that facilitates the generation of new NOD-based mouse models with the aim to improve the existing xenograft models. |
| Databáze: | OpenAIRE |
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