Yang Eric J Y, Nemhauser Jennifer L
Department of Biology, University of Washington, Seattle, WA, USA.
Quant Plant Biol. 2023 Oct 19;4:e12. doi: 10.1017/qpb.2023.10. eCollection 2023.
To support the increasingly complex circuits needed for plant synthetic biology applications, additional constitutive promoters are essential. Reusing promoter parts can lead to difficulty in cloning, increased heterogeneity between transformants, transgene silencing and trait instability. We have developed a pipeline to identify genes that have stable expression across a wide range of tissues at different developmental stages and have identified a number of promoters that are well expressed in both transient () and stable () transformation assays. We have also introduced two genome-orthogonal gRNA target sites in a subset of the screened promoters, converting them into NOR logic gates. The work here establishes a pipeline to screen for additional constitutive promoters and can form the basis of constructing more complex information processing circuits in the future.
为了支持植物合成生物学应用所需的日益复杂的电路，额外的组成型启动子至关重要。重复使用启动子部分可能会导致克隆困难、转化体之间的异质性增加、转基因沉默和性状不稳定。我们已经开发了一种流程来鉴定在不同发育阶段的广泛组织中具有稳定表达的基因，并鉴定了一些在瞬时（）和稳定（）转化试验中均能良好表达的启动子。我们还在一部分筛选出的启动子中引入了两个基因组正交gRNA靶位点，将它们转化为或非逻辑门。此处的工作建立了一个筛选额外组成型启动子的流程，并可为未来构建更复杂的信息处理电路奠定基础。
