Efficient genetic manipulation of the NOD-Rag1-/-IL2RgammaC-null mouse by combining in vitro fertilization and CRISPR/Cas9 technology.
Sci Rep
; 4: 5290, 2014 Jun 17.
Article
en En
| MEDLINE
| ID: mdl-24936832
ABSTRACT
Humanized mouse models have become increasingly important and widely used in modeling human diseases in biomedical research. Immunodeficient mice such as NOD-Rag1-/-IL2RgammaC-null (NRG) or NOD-SCID-IL2RgammaC-null (NSG) mice are critical for efficient engraftment of human cells or tissues. However, their genetic modification remains challenging due to a lack of embryonic stem cells and difficulty in the collection of timed embryos after superovulation. Here, we report the generation of gene knockout NRG mice by combining in vitro fertilization (IVF) and CRISPR/Cas9 technology. Sufficient numbers of fertilized embryos were produced through IVF, and a high rate of Fah gene targeting was achieved with microinjection of Cas9 mRNA, gRNA and single strand oligonucleotide DNA (ssDNA) into the embryos. The technology paves the way to construct NRG or NSG mutant mice to facilitate new humanized mouse models. The technology can also be readily adapted to introduce mutations in other species such as swine and non-human primates.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Técnicas In Vitro
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Técnicas Genéticas
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Proteínas Asociadas a CRISPR
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Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas
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Sistemas CRISPR-Cas
Límite:
Animals
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Female
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Humans
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Male
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Pregnancy
Idioma:
En
Revista:
Sci Rep
Año:
2014
Tipo del documento:
Article
País de afiliación:
Estados Unidos