Long-chain non-coding RNAs (lncRNAs) are a mysterious molecule that is about 200 nucleotides in length but does not encode any protein. Scientists have long wanted to know what such molecules in the genome do.
On December 15th, Science magazine published a new study found that researchers from the University of California, San Francisco identified 499 new lncRNAs, which laid the foundation for understanding the function of these small molecules. At the same time, the researchers also found that lncRNAs differ from most of the coding genes, which can act on a variety of cell lines, but surprisingly, 90% of the discovered lncRNAs affect only one cell type.
"Most cells have a lot of mysterious molecules like lncRNAs, but we don't know how many of them are specific and which are important for basic biological functions. This latest achievement surprised us that most lncRNAs are only one kind. The cell type is functional, and the extreme specificity of this RNA function has never been discovered before," said Daniel Lim, a neurologist and clinician at the University of California, San Francisco.
Previous studies have shown that certain lncRNAs play a key role in cell growth and function, as well as in the pathogenesis of the disease. Ten years ago, scientists discovered that a large part of the human genome was transcribed into lncRNAs, and some research groups pointed out dozens of lncRNAs. It is important for cells and even the body. However, only a few studies have discovered the function of lncRNAs, which is limited by research tools that analyze lncRNAs function and role cell types.
In the latest study, researchers used a very popular new tool: CRISPR technology to develop a CRISPR Interference Platform (CRISPRi) that targets 16,401 lncRNAs in seven different cell lines. The cell line includes six transformed cell lines, and a human induced pluripotent stem cell line. Using the CRISPRi tool, the researchers suppressed each candidate lncRNA, which screened thousands of sites and found the sites necessary for robust growth of 499 cells, 89% of which acted only on one cell type.
"The power of this technology comes from CRISPRi, which helps us turn off transcription of these non-coding transcripts," Lim said. In order to ensure the rapid screening of CRISPRi, the researchers also prepared 10 targeting RNAs for each locus to construct a library containing approximately 170,000 targeting RNAs. The researchers named the library CRiNCL (CRISPRi noncoding library) ), provided by Addgene.
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