Artificial mRNA

The Genetic Code Was Cracked Using Artificial mRNA Templates... 

Warthmann, N., H. Chen, S. Ossowoski, D. Weigel, and P. Herve. 2008. Highly specific gene silencing by artificial mRNAs in rice. PLoS ONE Mar 19 3(3) el829. 

After chemical synthesis of the mRNA sequence followed by pyrophosphorylation at the 5 end, the preparation of the artificial mRNA was completed by the enzymatic addition of a 5 cap and a 3 poly(A) tail. The synthetic strategy (Fig. 17) involved ... 

To test the ability of the artificial mRNA to support GLP-1 expression in whole cells, Chinese hamster ovary cells were transfected with artificial mRNA by electroporation. Twelve hours after transfection, cells were harvested and GLP-1 was determined by sandwich ELISA. mRNA derived from both the 130-nt and the 170-nt RNA supported GLP-1 expression, albeit weakly, whereas control RNA with no cap 1 or poly(A) tail did not. 

Fig. 19 GLP-1 expression of artificial mRNA in vitro. Artificial mRNA was added to a wheat germ cell-free protein synthesis system, and GLP-1 expression determined by sandwich ELISA. Each value is the average of three independent experiments, and the error bars indicate the standard deviation [129]...

Nagata S, HamasaM T, Uetake K et al (2010) Synthesis and biological activity of artificial mRNA prepared with novel phosphorylating reagents. Nucleic Acids Res 38 7845-7857... 

If DNA from two different species are mixed, denatured, and allowed to cool slowly so that reannealing can occur, artificial hybrid duplexes may form, provided the DNA from one species is similar in nucleotide sequence to the DNA of the other. The degree of hybridization is a measure of the sequence similarity or relatedness between the two species. Depending on the conditions of the experiment, about 25% of the DNA from a human forms hybrids with mouse DNA, implying that some of the nucleotide sequences (genes) in humans are very similar to those in mice. Mixed RNA DNA hybrids can be created in vitro if single-stranded DNA is allowed to anneal with RNA copies of itself, such as those formed when genes are transcribed into mRNA molecules. 

Reverse transcription is the copying of an RNA molecule back into its DNA complement. The enzymes that perform this function are called reverse transcriptases. Reverse transcription is used naturally by retroviruses to insert themselves into an organism s genome. Artificially induced reverse transcription is a useful technique for translating unstable messenger RNA (mRNA) molecules into stable cDNA. 

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