Journal
METHODS
Volume 191, Issue -, Pages 87-94Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2020.07.005
Keywords
Recombination; Transgenesis; Delivery; loxP; Conditional; MEF; EC
Funding
- Czech Health Research Council [17-32727A]
- CAS [RVO: 86652036, RVO 68378050]
- Ministry of Education, Youth and Sports (MEYS) [LQ1604, LM2015040, LM2018126]
- European Regional Development Fund [CZ.1.05/1.1.00/02.0109, CZ.1.05/2.1.00/19.0395]
- MEYS
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The Cre-loxP recombination system is a powerful tool for genome engineering, with one application being the induction of Cre recombination in genetic mouse models. A technique of delivering Cre protein into mouse zygotes by electroporation is described as simple, affordable, and highly efficient, resulting in faster generation of desired models. Additionally, it is shown that the method is suitable for allelic conversion in primary cells derived from conditional mouse models.
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.
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