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214 || AWSAR Awarded Popular Science Stories - 2019
sprouting from a certain kind of vessels with a basket-like structure called sub-intestinal vessels. When we introduced excess ELNC, we could see extra vessels sprouting off from the basket. This hinted that in normal conditions ELNC is required for routine blood vessel maintenance.
All information in a cell is written in a molecule called deoxyribonucleic acid or DNA. The information or “message” that any RNA holds is “copied” from DNA. Any change in DNA can alter the RNA as well. We used a technique called gene-editing to alter the DNA containing ELNC. Here we aimed to have a complete abolishment of ELNC function or a “knockout”, opposed to suppression by knockdown technique. Using microinjection, we introduced artificially synthesized gene- editing molecules into one-cell stage, double- labeled zebrafish. We could see hemorrhages in the head region at ages two days and above in these fishes. This observation
the compartment which is like the control centre of a cell, called nucleus. The other fraction contained all other compartments, called cytoplasm. The nucleus was further divided in two fractions: one containing DNA called chromatin, and the other nucleoplasm. We found that ELNC is localized in the chromatin: this told us that ELNC could be involved in processes that happen around DNA or formation of RNA from it.
When we first chose ELNC for our study, we had a hunch that it could be regulating a protein already known to have a function in endothelial cells. This was because in DNA ELNC lies near the DNA stretch that has the information to make this protein, say Protein-E. So in our ELNC knockdown and knockout zebrafish systems showing hemorrhage we started investigating how this Protein-E was affected. While Protein-E itself isn’t affected, we have a hint that ELNC could be regulating the
process which generates RNA required to make Protein-E.
Meanwhile, we also wondered if ELNC could be a player in endothelial function in humans. The incredible thing about the zebrafish model is the similarity in DNA between zebrafish and humans, almost 70%! That is, 7 out of 10 times we identify a new protein or RNA in zebrafish we could apply that knowledge to humans. But here’s the tricky part: lncRNA are known to have entirely varying DNA make- ups in different organisms. Yet there is a relatively easy way to find human ELNC. If we could speculate a certain lncRNA in humans could be the counterpart of zebrafish
lncRNA be able to do the same function. This
backed our results from the knockdown experiment. We further raised these fishes with altered ELNC DNA and generated progeny, which also displayed hemorrhage. This was exciting, because by independent techniques we could show that disruption of ELNC could affect blood vessel architecture or integrity and even lead to hemorrhage!
Now that we knew ELNC was important for endothelial cell function, we wanted to know what exactly the lncRNA did in the cell. A cell is divided in many compartments, each of which perform distinct actions, all hand-in-hand for the survival of the cell. We isolated two fractions of the cell mixture: one
that contained
All information in a cell is written in a molecule called deoxyribonucleic acid or DNA. The information or “message” that any RNA holds is “copied” from DNA. Any change in DNA can alter the RNA as well. We used a technique called gene-editing to alter the DNA containing ELNC. Here we aimed to have a complete abolishment of ELNC function or a “knockout”, opposed to suppression by knockdown technique.