RNA-based probes: Spinach aptamers

In course

Description

This lecture delves into the engineering and application of RNA-based probes, focusing on Spinach aptamers as GFP-mimics of RNAs. Starting with a refresher on GFP chromophore maturation, it progresses to discussing nucleic acid aptamers, SELEX technology, and the evolution of Spinach as a GFP-mimic of RNA. The lecture explores in vitro transcription, cDNA, aptamer selection, and characterization, emphasizing the critical evaluation of the manuscript. It also covers the tracking of RNA with Spinach, comparing it with GFP fusion protein design. The session concludes with a critique of experimental data and movies related to RNA localization and export, highlighting the key learning outcomes and applications of RNA-based probes.

Official source

https://mediaspace.epfl.ch/media/0_q5myustl

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Ribonucleic acid (RNA) is a polymeric molecule that is essential for most biological functions, either by performing the function itself (Non-coding RNA) or by forming a template for production of proteins (messenger RNA). RNA and deoxyribonucleic acid (DNA) are nucleic acids. The nucleic acids constitute one of the four major macromolecules essential for all known forms of life. RNA is assembled as a chain of nucleotides.

Threose nucleic acid

Threose nucleic acid (TNA) is an artificial genetic polymer in which the natural five-carbon ribose sugar found in RNA has been replaced by an unnatural four-carbon threose sugar. Invented by Albert Eschenmoser as part of his quest to explore the chemical etiology of RNA, TNA has become an important synthetic genetic polymer (XNA) due to its ability to efficiently base pair with complementary sequences of DNA and RNA. However, unlike DNA and RNA, TNA is completely refractory to nuclease digestion, making it a promising nucleic acid analog for therapeutic and diagnostic applications.

Antisense RNA

Antisense RNA (asRNA), also referred to as antisense transcript, natural antisense transcript (NAT) or antisense oligonucleotide, is a single stranded RNA that is complementary to a protein coding messenger RNA (mRNA) with which it hybridizes, and thereby blocks its translation into protein. The asRNAs (which occur naturally) have been found in both prokaryotes and eukaryotes, and can be classified into short (200 nucleotides) non-coding RNAs (ncRNAs).

RNA editing

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