Department of Pharmacology, Genome Sciences, and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, USA.
Department of Biochemistry, Genome Sciences, and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, USA.
Trends Genet. 2018 May;34(5):333-340. doi: 10.1016/j.tig.2017.12.008. Epub 2018 Jan 11.
The Cre-lox recombination approach is commonly used to generate cell-specific gene inactivation (or activation). We have noticed that the breeding and genotyping sections of papers utilizing Cre-lox techniques are frequently incomplete. While seemingly straightforward, there are important considerations that need to be implemented in the breeding and genotyping methods to prevent the introduction of experimental confounds. Germline recombination and transient expression of Cre recombinase during development are some examples of the complications that can occur, and conventional genotyping methods may fail to identify these events. In this opinion article, we highlight the importance of testing for unexpected recombination events, suggest strategies to isolate and minimize adverse recombination events, and encourage editors and reviewers to expect more definitive statements regarding the validation of genotyping.
Cre-lox 重组方法常用于生成细胞特异性基因失活（或激活）。我们注意到，利用 Cre-lox 技术的论文的繁殖和基因分型部分经常不完整。虽然看似简单，但在繁殖和基因分型方法中需要考虑一些重要的因素，以防止引入实验混淆。生殖系重组和 Cre 重组酶在发育过程中的瞬时表达就是可能发生的一些复杂情况的例子，传统的基因分型方法可能无法识别这些事件。在这篇观点文章中，我们强调了测试意外重组事件的重要性，提出了分离和最小化不利重组事件的策略，并鼓励编辑和审稿人对基因分型的验证提出更明确的声明。
