RNAi mechanism | RNA interference pathway using siRNA and shRNA

welcome back friends after a long time I'm making videos and in this video we'll be talking about RNA interference on RNA I now getting I'm getting many requests from different viewers to make videos on RNA I si RNA shrna mi RNA now this type of things the total total are an interference thing it is very very important and it's a very hot topic of study nowadays so what is are any interference now there are a lot of complications if you search for images you will find many fascinating images complicated images because you know it's very true fact we all understand DNA biology very well but very few of us actually care about RNA but this is the time for caring about RNA because RNA biology is equally being important in different diseases in cancer and also you know in different aspects of our life so the first thing that we are going to talk about today is about the RNA I on RNA interference it's also called as a RNA interference or interfering RNA so let me write it here RNA interference we need to understand what is RNA interference now are any interference is a phenomena inside the cell which targeted ly select mRNA strand and chop them up right it's a whole process inside eukaryotic cells or prokaryotic cell because we found the evidence of RNA I in bacteria we have found it in fungi algae as well as plants and animals it is very much useful in plant biology why we will studying later now are any difference that have told you that they targeted li the selected target and that is sequential selection of a particular target and they break down that target you break down that target now the target here by RNA interference our common protein coding RNAs which is also known as mRNA so RNA interference actually target mRNA and break it down so that there is no protein produced from the mRNA right so that is the actual goal now the basic idea is the same generally RNA is a single stranded molecule we all know that right it should have a single strand and the base pairs everything normal basis not pair everything is fine so whatever is there inside a cell if there is by any chance they produce a double stranded RNA that double stranded RNA can be very very dangerous because the double stranded RNA could be a very specific target site to produce this RNA interference and to ultimately break down the actual protein coding RNA or mRNA right because the fundamental law of producing an interference or RNAi things like shrna or mi RNA or si RNA are the double stranded RNA inside the cell so if there is any production of double stranded RNA inside the cell these things can lead to the production of si RNA it can produce is H RNA it can also produce a my RNA right so what are the full forms of all this essay RNA means small interfering RNA shrna is short hairpin RNA mi RN is simply micro RNA right so let me write RN is here so these are the short forms of these three RNAs right so all of this type of RNA si RNA shrna mi RNA they are produced from double-stranded RNA so if by any means inside the cell a double-stranded RNA is generated in that case they can have the potential to produce all this kind of are an inside and if they produce this type of RNA inside a cell these things can actually lead to the silencing silencing of the mRNA the actual protein coding RNA and that is called RNA mediated gene silencing and they bring out this gene silencing process by combining many other proteins together and that protein complex is called as RI SC RNA interference silencing complex or RI SC so if they produce any of this they will form our ISC early interfering silencing complex now this risk factor with this si are NR shRNA will ultimately silence the mRNA that is the actual protein coding RNA it will break down this error how they can silence it will actually degrade that on break that RNA down smaller fragments so as a result protein synthesis will not be possible and they can't produce any required protein inside the cell that is the target process of RNA interference right now once we know this risk is required so to produce the risk they require certain enzymes and other molecules to be associated actually the they have association of certain enzymes called slicer it's a protein right usually it is required and it was found inside C elegans sinner of deities elegans and in their body we have seen several effects of our interference and we have seen slicer to be a part of risk in C elegans similarly we can also see another very important protein that is as Argonaut Argonaut is another protein which is also commonly found in plants as well as animals in the RNA interference system to be a part of risk okay now what are this molecule slicer but all above there is another protein which is the mother to produce this as high RNA shRNA an mi RNA from double-stranded RNA that protein is termed as Dicer very very important protein with respect to the RNA interfering system because this geyser should be active to produce any one of this si RNA shrna or mi RNA from the micro double stranded RNA inside the cell right this Dicer is nothing but RNA is free type of enzyme now what we mean by RNA is RNA is 3 what we mean by our NST is this type of enzymes RNA 3 type of enzymes they are double-stranded RNA specific endonuclease that means they will specifically bind to double-stranded RNA and it will break down the RNA from the middle in the nucleus not from the terminal if it breaks down the RNA from the terminal it should be called EXO nucleus but it is breaking down from the middle so endo nucleus right so Dicer is always active there and the slicer here which you talked about and are the no argument is also our nh3 type of enzyme there slicer in other hand is RNA is H type of enzyme over there right so they all can actually function accordingly but the major task is for the Dicer because the Dicer ultimately at the first produce this is high RNA mRNA or shrna from the double stranded RNA and once the production is complete then this type of small interfering RNA particles will go and it will form RISC complex with other proteins called slicer or Argonaut or any other proteins are there we'll be talking about them later in the future videos now once they produce that risk complex they will go and they target they select their target right because you know this is very very specific that is another important property this whole process is specific that means it is specified that which part of the mRNA should be degraded or which among knee should be degraded right because you know it's all about pairing with pieces we have RNA which is a single-stranded that RNA can form bond with itself and that can form hairpin and imagine similarly that RNA once it is degraded with slicer and are not in the RISC complex will ultimately bind with certain sequences in the mRNA only so that RNA it depends on which sequence they can bind right so suppose it's a 200 base pairs long mRNA sequence right 200 bases long mRNA now that Dicer is that that whole sh are any odds let's say the whole si RNA can actually bind with the sequence number 50 to 65 and in that case they will select that sequence and then cut it from let us say a 67 number base pair position and the RNA is now broken down into two and you imagine the situation that's how the whole process work it is very specific specifically select the mRNA attached with it break it down right so that is called as the RNA I or RNA interference this is the simplest explanation I could have given you about RNA interference it is a huge complicated topic but believe me what you need to understand in the basic way so that you can explain it to someone with a layman in biology this is not an interference this is this task now what are the advantages of studying or an interference because you know RNA interference is a hugely important system because we can silence a gene inside we are not targeting the DNA the DNA is not hampered the code the complete blueprint is not hampered but what we can achieve from here we can block the synthesis of the protein we can block the translation we can break down that RNA so by using this process we can actually achieve miracles so what this this is hi this complete set of RNA interference is actually everywhere it's been discovered later lately but actually it is there from the earlier times it is acting all the time right I've told you people don't have a very good arch to understand biology in RNA biology very well they always start to talk about DNA protein synthesis but very few about RNA and this is very very important guys because you know suppose in a disease let's say this is a cell this is a nucleus and what happens in a particular disease that nucleus they produce a gene they produce an mRNA from the Dame RNA they're producing certain proteins that protein is causing the cell to die or causing a particular disease now if we identify that mRNA and we can specifically design an SI RNA which will targeted Lee cleave that mRNA which produces the malfunctioning protein inside so let's say we we build that si RNA we inject the si RNA inside that cell so that that is high RNA will act and bind with this this mRNA and degrade the mRNAs which produces the malfunctioning protein so ultimately what happens all the proteins will be completely taken care of so there will be no malfunctioning protein inside the cell anymore and the cell will start functioning normally so you can see the possibility is huge but there is a question that as we are preventing this in the translation level right for that reason we need to administer this quite often because you know suppose we add this si RNA and we block it we break down the mRNA sin is fine now after certain time in the next time the gene will express it will Express the mRNA again in that case we need to consider and take this si RNA again from outside so that is the problem we need to continuously administer that si RNA or whatever medication we are in to control those type of diseases but there is a salut for that too and the solution here is to not only take this SI RNA from there directly what we do we instead inject the cell with a vector and that vector itself produces a type of double-stranded RNA so if that vector will continue continuously Express with the chromosomes of that cell so it will produce double-stranded RNA continuously and from the double-stranded RNA inside the cell they easily produce si RNA or mi RNA which will ultimately break down the targeted mRNA there so you can see the possibilities are limitless if you understand RNA biology very well right so that's all about the overview of RNA I from the next videos we'll be making actually four videos in this whole topic this is the first video in the last three videos or we'll be talking about you know si RNA shRNA and mi RNA sequentially so let's talk about it in the future videos stay tuned watch it don't forget to subscribe and go to my website see videos and see other links there thank you

Channel: Shomu's Biology

Views: 281,952

Rating: 4.8880997 out of 5

Keywords: suman bhattacharjee, shomus biology, biology online, biology courses, life science, biology major, RNA Interference (Award-Winning Work), Small Interfering RNA, Small Hairpin RNA, rnai, rna interference, sirna, shrna, mirna, rnai mechanism, rnai pathway, rnai transfection, rnai therapeutics, microrna, micro rna, gene silencing

Id: mdwzpKTxoUI

Length: 14min 33sec (873 seconds)

Published: Sat Feb 21 2015