Sequencing of yeast RNAi libraries for improved protein production
Ontology highlight
ABSTRACT:
PROVIDER: PRJEB27502 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| ERR2672940_1.fastq.gz | Fastqsanger.gz | |||
| ERR2672940_2.fastq.gz | Fastqsanger.gz | |||
| ERR2672941_1.fastq.gz | Fastqsanger.gz | |||
| ERR2672941_2.fastq.gz | Fastqsanger.gz | |||
| ERR2672938_1.fastq.gz | Fastqsanger.gz |
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Project description:Improvement and research of plants depends on the long-term expression of transgenes. However, the durability of transgene expression is routinely hampered by silencing pathways that start as the post-transcriptional process of transcript cleavage by RNA interference (RNAi). To avoid transgene silencing, we aimed to inhibit the sorting of transgene mRNAs into RNAi. We manipulated a well-studied protein / RNA-binding module from Arabidopsis into a transgene transcript, where the transcript is now bound by an engineered RNA-binding protein that preferentially sorts the RNA into translation, rather than RNAi. We used the Cas9 transcript as a proof-of-principle and demonstrated higher Cas9 protein production and editing rates. In addition, transgenes with the engineered protein / RNA-binding module had improved long-term durability of transgene expression, as after four generations these plants had higher Cas9 protein accumulation and lower levels of DNA methylation, a hallmark of transgene silencing. Our engineered system represents a successful manipulation of post-transcriptional RNA sorting for improved transgene performance, and could be applied to any transgene transcript.
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