Efficient allele conversion in mouse zygotes and primary cells based on electroporation of Cre protein.
Conditional
Delivery
EC
MEF
Recombination
Transgenesis
loxP
Journal
Methods (San Diego, Calif.)
ISSN: 1095-9130
Titre abrégé: Methods
Pays: United States
ID NLM: 9426302
Informations de publication
Date de publication:
07 2021
07 2021
Historique:
received:
08
04
2020
revised:
23
06
2020
accepted:
16
07
2020
pubmed:
28
7
2020
medline:
5
1
2022
entrez:
28
7
2020
Statut:
ppublish
Résumé
Cre-loxP recombination system is a powerful tool for genome engineering. One of its applications is found in genetic mouse models that often require to induce Cre recombination in preimplantation embryos. Here, we describe a technically simple, affordable and highly efficient protocol for Cre protein delivery into mouse zygotes by electroporation. We show that electroporation based delivery of Cre has no negative impact on embryo survival and the method can be easily combined with in vitro fertilization resulting in a significantly faster generation of desired models. Lastly, we demonstrate that Cre protein electroporation is suitable for allelic conversion in primary cells derived from conditional mouse models.
Identifiants
pubmed: 32717290
pii: S1046-2023(20)30114-6
doi: 10.1016/j.ymeth.2020.07.005
pii:
doi:
Substances chimiques
Cre recombinase
EC 2.7.7.-
Integrases
EC 2.7.7.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
87-94Informations de copyright
Copyright © 2020. Published by Elsevier Inc.