School of Biological Engineering, Dalian Polytechnic University, Dalian 116034, PR China.
Laboratory of Biotechnology, Dalian Institute of Chemical Physics, CAS, 457 Zhongshan Road, Dalian 116023, PR China.
FEMS Yeast Res. 2019 May 1;19(3). doi: 10.1093/femsyr/foz031.
The red yeast Rhodosporidium toruloides is an excellent microbial host for production of carotenoids, neutral lipids and valuable enzymes. In recent years, genetic tools for gene expression and gene disruption have been developed for this red yeast. However, methods remain limited in terms of fine-tuning gene expression. In this study, we first demonstrated successful implementation of RNA interference (RNAi) in R. toruloides NP11, which was applied to down-regulate the expression of autophagy related gene 8 (ATG8), and fatty acid synthase genes (FAS1 and FAS2), respectively. Compared with the control strain, RNAi-engineered strains showed a silencing efficiency ranging from 11% to 92%. The RNAi approach described here ensures selective inhibition of the target gene expression, and should expand our capacity in the genetic manipulation of R. toruloides for both fundamental research and advanced cell factory development.
红色酵母鲁氏酵母（Rhodosporidium toruloides）是生产类胡萝卜素、中性脂肪和有价值的酶的绝佳微生物宿主。近年来，已经为这种红色酵母开发了用于基因表达和基因敲除的遗传工具。然而，在精细调控基因表达方面，方法仍然有限。在本研究中，我们首先证明了 RNA 干扰 (RNAi) 在 R. toruloides NP11 中的成功实施，该方法分别用于下调自噬相关基因 8 (ATG8) 和脂肪酸合酶基因 (FAS1 和 FAS2) 的表达。与对照菌株相比，RNAi 工程菌株的沉默效率范围为 11%至 92%。本文所述的 RNAi 方法可确保靶基因表达的选择性抑制，应扩大我们在鲁氏酵母遗传操作方面的能力，既用于基础研究，也用于先进的细胞工厂开发。
