Shanghai NCRNA sequencing experiment

Shanghai NCRNA sequencing experiment

RNA, LNCRNA) refers to an RNA molecule that does not have protein encoding function between 200 to 100,000 NT. They have two forms with Polya tails and do not bring PolyA, with low-conservative, tissue specific expression and low abundance. The type of lncRNA far exceeds the encoding RNA, and the sequence of 4% to 9% of the mammaline genome sequence produces LNCRNA transcripts, while the proportion of corresponding protein encoded RNA is only 1% to 5%. The current research suggests that LNCRNA's functional mechanism mainly includes participating in transcriptional regulation, transcriptional regulation, protein transport and mRNA degradation. LNCRNA has become a hot spot in the field of non-encoded RNA research, with high-throughput sequencing, researchers can quickly obtain LNCRNA associated with disease or specific biological processes and conduct in-depth studies.

Data Analysis Technology Route:

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New LNCRNA prediction results (Shanghai NCRNA sequencing experiment )

Sample requirements 1) Sample type: cell, fresh tissue or RNA sample.
2) Sample quantity: Cell samples provide at least 1 u0026 Times; 107 cells, tissue samples provide at least 300 mg tissue blocks or slices, RNA samples, please provide 10 u0026 mu; g above total RNA.
3) Sample quality: RNA has no obvious degradation, the extraction total RNA OD260 / 280 value is between 1.8 ~ 2.2, concentration u0026 ge; 500 ng / u0026 mu; L, 28S: 18S u0026 ge; 1.5, RIN u0026 GE; Sign in.

4) Sample preservation: Cell samples or fresh tissue blocks (cut to 50 mg of small pieces) can be stored with Trizol or RNA protective agents or liquid nitrogen survival, -80 ° C. RNA samples are soluble in ultrapure water of ethanol or RNA-Free, and -80 ° C is stored. During the storage period, avoid repeated freeze-thaw.

5) Sample Transport: Samples are placed in 1.5 ml tubes, sealing the membranes, dry ice transport. References 123] [1] The GENCODE v7 catalog of human long noncoding RNAs: Analysis of their gene structure, evolution, and expression Genome Res 2012 22:... 1775-1789 [2] A long noncoding RNA regulates photoperiod-sensitive Male Sterility, An Essential Component of Hybrid Rice. PNAS, 2012, 109: 2654-2659

LONG NONCODING RNAS with Snorna Ends. Molecular Cell, 2012, 48: 219 u0026 ndash; 230 Using NGS method: A type of biplasid contains small nucleus RNA (snorna) in the splicing A type of new type of long-coded RNA can be formed in the process, named SNO-lncrnas. There are five SNO-lNCRNAs in the tightly associated region of human disease Prader-Willi SYNDROME, which have extremely high expression in human embryonic stem cells and abnormal stable. Functional studies have shown that these SNO-LNCRNAS processes are collected near their transcription sites to form a new cell nuclei (which also contains a specific binding site of multiple splice regulators FOX2 proteins, which can be like u0026 ldquo Sponge u0026 rdquo; as the FOX2 in the cell nucleus, regulates the distribution of FOX2 in the cellular nucleus, which in turn affects the selective splicing regulation of FOX2 on specific mRNA substrates. These SNO-lNCRNAs were missing in PWS syndrome, which suggesting that these new RNA molecules may be associated with pathologies of PWS syndrome.

Research Results

1. The candidate Sno-lNCRNA is found using RNA sequencing, and verify confirmation.

2. The regulation mechanism of SnO-lNCRNA was found by subsequent test results.