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Mangoes are naturally rich in Vitamin A, B6, C and E, calcium, iron, magnesium, potassium, protein, sugar and fibre. The antioxidant compounds in mango protect you from cellular damage and many types of cancers. With high levels of pectin and fibre, mango lowers cholesterol and blood pressure. Mango also restores the body?s alkaline-acid balance

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From The Earth is a fully licensed cannabis dispensary built with our community in mind. Our team personalizes each customer experience based on your unique needs. We are here to help you learn about cannabis, live with the benefits, and grow as an individual.

The sativa-leaning strain Ghost Lime Pop is defined by its ripe orange bouquet accented with a touch of earthy-pine funk from its predecessor Ghost OG. This single-source live resin is cultivated by the legacy growers at Sonoran Roots.

RNA molecules play vital roles in many cellular processes. Visualising their dynamics in live cells at single-molecule resolution is essential to elucidate their role in RNA metabolism. RNA aptamers, such as Spinach and Mango, have recently emerged as a powerful background-free technology for live-cell RNA imaging due to their fluorogenic properties upon ligand binding. Here, we report a novel array of Mango II aptamers for RNA imaging in live and fixed cells with high contrast and single-molecule sensitivity. Direct comparison of Mango II and MS2-tdMCP-mCherry dual-labelled mRNAs show marked improvements in signal to noise ratio using the fluorogenic Mango aptamers. Using both coding (-actin mRNA) and long non-coding (NEAT1) RNAs, we show that the Mango array does not affect cellular localisation. Additionally, we can track single mRNAs for extended time periods, likely due to bleached fluorophore replacement. This property makes the arrays readily compatible with structured illumination super-resolution microscopy.

Since their development, fluorogenic RNA aptamers have been poised as a potential complementary alternative to the MS2 system15,16,17,18,19,20,21,22,23,24. Their fluorogenicity enables the induction of fluorescence upon RNA expression and can facilitate high-contrast imaging and offer the potential to use shorter RNA tags. These RNA aptamers after being transcribed, induce fluorescence by binding a fluorogenic dye making it thousands of times brighter upon binding25. A wide range of fluorogenic RNA aptamers have been selected and shown to induce aptamer specific fluorescence in live cells. Two of the most promising families of small monomeric aptamers are Spinach and Mango17,18,19. It has long been thought that aptamer arrays, similar to the MS2 cassettes, could enable single-molecule imaging with improved sensitivity. However, both the folding and fluorescence stability of the Spinach aptamer have hindered high-resolution imaging of such arrays20. More recently, arrays of the Pepper aptamer have been used to image RNAs with improved resolution26, but the use of fluorescently labelled proteins is still required for single-molecule detection27. The brightness, thermodynamic stability and high-affinity of Mango aptamers bodes well to the accurate detection of RNAs abundance in a cellular environment. The recent isolation of brighter and more photostable Mango aptamers, as well as the detection of as low as five tagged RNAs within an individual foci, suggest the potential to image single molecules of RNA using Mango arrays28. Here we show the development of stably folding Mango II repeats and their ability to directly image both coding and non-coding RNAs at single-molecule resolution without affecting their known localisation patterns. By making dual Mango and MS2 arrays of exactly the same size we directly compare the imaging capabilities of the MS2 and Mango systems in a head to head fashion. Furthermore, exchange of the Mango fluorogenic dye in fixed samples extends imaging times, which also benefits super-resolution techniques such as structured illumination microscopy (SIM).

Since the development of fluorescently tagged bacteriophage coat protein to image RNA molecules in cells10, single-molecule imaging of mRNAs has provided an important window into the dynamic life of RNA molecules during transcription, splicing, post-transcriptional modification, translation and degradation2,5,6,7,47,48,49. However, the stability of the capsid bound RNA tag within rapidly degraded mRNAs in yeast has highlighted the need to develop alternative labelling strategies to improve both imaging contrast and reduce aberrant degradation products. In addition to the recently modified MS2-SL sequences13, fluorogenic RNA aptamers represent an interesting and complementary alternative approach. Fluorogenic RNA aptamers give the added benefit of producing a specific fluorescent signal only in the presence of tagged RNA. To date, fluorogenic RNA aptamers have been used to image short non-coding RNAs with attempts at improving the resolution of imaging with tandem aptamer arrays20. However, the fluorescent stability of these arrays both in vitro and in bacterial cells has hindered the observation of single molecules. Recently, two new aptamer-based RNA imaging technologies have emerged both naming their aptamers Pepper. The first using RNA-stabilised fluorescent proteins able to image single mRNAs in live cells27. The second, is a fluorogenic RNA aptamer that has been shown to enable RNA imaging in cells, but not at the single molecule level26. RNA Mango is likely compatible with these technologies while enabling direct RNA imaging at single molecule resolution without the need for fluorescent proteins.

While the long Mango arrays used in this study allowed us to directly compare with existing MS2 arrays in live cells, there appears to be no fundamental reason to use such long repeat lengths with the Mango system. In this and previous studies28 we have routinely been able to detect low Mango copy number in cells suggesting that array size minimisation is likely. As widefield microscopy uses relatively intense laser light to image live cells, the photobleaching conditions were quite harsh. Further improvements in live-cell imaging methodology are therefore easily imagined as a consequence. Specifically, further optimisation of fluorophore concentration and Mango array size could allow continuous RNA imaging via fluorophore exchange within living cells. This combined with the use of increasingly standard microscope imaging methodologies such as light sheet51,52, orbital tracking53, laser pulse sequence, or laser scanning methodologies54 appear highly likely to further improve this already compelling approach to the routine imaging of single RNAs within live cells via the use of RNA Mango.

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