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      "body": "I do CRISPR every day and have made dozens of genetically modified cell lines. I can easily do this, but your article is overly simplified. Having said that, I haven't done any reading on this disease or the mutation that causes it, but I'm certain I could fix it... And that you wouldn't want to use PEI for the transfection ;)",
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      "body": "Antibiotics work on bacteria, so viruses are not effected by antibiotics. Do not take antibiotics for a viral infection.\n\nNow that that is out of the way, I like the idea of using bacteriophages to take out antibiotic resistant strains of bacteria. A bacteriophage is a virus that specifically can target and lyse (kill) the target bacteria. Of course, within a population of bacteria, there can be naturally occurring resistance to phages. This resistance, of course, can be due to the bacterial immune system known as CRISPR. Funny how that works.",
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      "body": "This paper has since been retracted. Their data was poorly obtained and incorrectly analyzed.\n\nhttps://www.nature.com/articles/nmeth.4664",
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      "body": "That's a good question, I'd have to ask my supervisor. He's been working on this for a while tho and what you suggested is the traditional way, there's probably a reason why we're not doing it like that. It's mice btw.",
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      "body": "@suesa why not flox the gene of interest you want to knockout, and do tissue specific Cre expression by throwing in an inducible Cre driven by a neuron specific promoter? I guess it depends on what model system you're using",
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      "title": "",
      "body": "Good to know! I'll be using it to create a loxp site in a specific gene of mice. I'll message you when it doesn't work :D",
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      "body": "Cool article, but the Cre/Loxp system will (most likely) never be used in a clinical setting for humans, at least not in the United States or anywhere with a decent regulatory body. Perhaps the biggest reason is that extra, non-native, sequences of DNA are being inserted into the genome (loxp sites). Even after removal with Cre, there is a \"scar\" that leaves behind one loxp site in the genome, which is considered a no-no.\n\nAs for wild-type CRISPR/Cas9 for the clinical setting, I think it inherently is dangerous to use because it can have a lot of unintended, off-target effects. My opinion is that the current state of CRISPR/Cas9 is good for quickly and cost-effectively investigating genetic modifications, but a more specific version or a new method of gene editing that is more specific would need to be used for high success rates, and low safety concerns, in a clinical setting.",
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      "body": "@schay and @suesa send me any CRISPR/Cas9 questions you have. I use it every day to make new human stem cell lines.",
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      "body": "Gene editing is a bit anti-climactic and is mostly imagination and your own knowledge.\n\nExample: Mix a drop of this colorless liquid with a drop of that colorless liquid. Put that colorless liquid into cells. Wait a week or so for the cells to divide so that you have more of them, then check if the cells got the desired genetic modifications. So it is important that you have the knowledge of what components are inside each of the previously mentioned colorless liquids and what they are doing, but otherwise it looks the same to the observer.\n\nI know this because I do it almost every day in human pluripotent stem cells.",
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      "title": "",
      "body": "I use Gibson assembly when I generate most plasmids, but sometimes you can't. But you'd be surprised how few people use Gibson assembly or even understand it. It's very useful and simple, so you'd think nearly everyone generating plasmids would use it...",
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      "body": "Good, nicely explained article :) Easy to follow",
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      "body": "Good, nicely explained article :) Easy to follow",
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      "permlink": "re-alphacoco-re-clausewitz-fascinating-genetics-cloning-vectors-the-plasmid-20180210t151750579z",
      "title": "",
      "body": "I think there is still too much controversy going around about who discovered CRISPR/Cas9 versus who figured out its best use. Additionally, the Nobel prize isn't awarded posthumously, so I think priority was given to some people who had amazing contributions to the field but are near the end of their life.. The CRISPR groups are all young :)",
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      "body": "Congratulations @fighton! You have completed some achievement on Steemit and have been rewarded with new badge(s) :\n\n[![](https://steemitimages.com/70x80/http://steemitboard.com/notifications/firstpost.png)](http://steemitboard.com/@fighton) You published your First Post\n\nClick on any badge to view your own Board of Honor on SteemitBoard.\nFor more information about SteemitBoard, click [here](https://steemit.com/@steemitboard)\n\nIf you no longer want to receive notifications, reply to this comment with the word `STOP`\n\n> By upvoting this notification, you can help all Steemit users. Learn how [here](https://steemit.com/steemitboard/@steemitboard/http-i-cubeupload-com-7ciqeo-png)!",
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      "title": "Scientists develop new therapeutic approach to ALS",
      "body": "Scientists at the University of Southern California have come up with a new approach to study disease mechanisms of amyotrophic lateral sclerosis (ALS, motor neurone disease, or Lou Gehrig's disease). The multinational team of scientists established an in vitro (in a dish) model system that involves directly changing a blood cell into a motor neuron, the main cell type that is killed in ALS. So, the team can take a blood cell from an ALS patient and turn it into a motor neuron to study the disease. The patient derived induced motor neurons actually die more rapidly in vitro than motor neurons derived from the blood of healthy control counterparts, thus recapitulating the disease and creating a model to study ALS in the lab.\n\nPerhaps of most notable scientific significance, this paper establishes a new model system for studying neurodegenerative diseases in the cell type effected by the disease. The group exhaustively started this model system, and screened thousands of different drug-like molecules on the cells to identify molecules that could help the diseased cells survive longer in the dish. These findings will hopefully translate to help patients who have the disease.\n\nInterestingly, many different mutations can cause ALS, but this paper proposes a mechanistic convergence in one cellular pathway that can be modulated by drugs identified in their drug screen. Very exciting research indeed!\n\nReferences: Shi, et al. Nature Medicine doi:10.1038/nm.4490",
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      "author": "carlgbush",
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      "title": "",
      "body": "Hi @fighton, thank you for taking time to comment, you are completely right about gene drive. In this article I only talk about the natural occurrences and mechanisms of homing endonuclease, that's what @kilbride question was about. I will talk about Cas9 and how this mechanism is used for gene drive in the next article :)",
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      "parent_permlink": "re-jepper-activate-genes-with-light-optogenetic-control-of-intracellular-signalling-20180203t174200052z",
      "author": "jepper",
      "permlink": "re-fighton-re-jepper-activate-genes-with-light-optogenetic-control-of-intracellular-signalling-20180203t194722144z",
      "title": "",
      "body": "Wow! It is great to have someone that actually uses the method here. Sounds extremly interesting. You should write an article about it!",
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      "parent_permlink": "activate-genes-with-light-optogenetic-control-of-intracellular-signalling",
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      "permlink": "re-jepper-activate-genes-with-light-optogenetic-control-of-intracellular-signalling-20180203t174200052z",
      "title": "",
      "body": "@@ -59,16 +59,35 @@\n eprogram\n+/transdifferentiate\n  human s\n",
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      "parent_permlink": "activate-genes-with-light-optogenetic-control-of-intracellular-signalling",
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      "permlink": "re-jepper-activate-genes-with-light-optogenetic-control-of-intracellular-signalling-20180203t174200052z",
      "title": "",
      "body": "@@ -60,16 +60,22 @@\n program \n+human \n skin cel\n",
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      "title": "",
      "body": "I used channelrhodpsin when I figured out a novel way to reprogram skin cells directly into skeletal muscle. I would shine blue light on the cells, causing the channelrhodopsin to open and depolarize the cell, causing the newly made muscle cell to contract in the dish. The functional readout allowed me to compare efficiencies across multiple conditions to figure out the best one! :)",
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      "parent_permlink": "re-kilbride-re-carlgbush-gene-drive-and-the-engineering-of-extinction-part-i-20180201t230934600z",
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      "title": "",
      "body": "Hi @kilbride and @carlgbush. I perform CRISPR/Cas9 gene editing in human pluripotent stem cells, so I am familiar with these topics. Yes, self damage and apoptosis (termination of the whole cell, \"programmed cell death\") can occur. Pretty much nothing in biology is 100%, so the desired editing events, particularly transmission of the gene of interest, is a fairly rare event. But, there are many things to consider when designing your gene targeting strategy. My understanding is that gene drive in its current form is more used to remove a gene from a population. Removing a gene is a much easier process than making a specific change because you are only relying on the repair machinery of the cell to accidentally repair the double stranded DNA break with an insertion or deletion (termed indel) of one or a few additional/less DNA bases, thus causing a frameshift mutation and essentially removing a gene product.\n\nI would think to actually integrate the \"homing endonuclease\" (really a Cas9 and a few guide RNAs targeting the region of the genome that you want) directly into the gene that you want to remove would be most efficient, particularly if you were to add a stop codon just downstream of your homing endonuclease. So if the repair process is through homologous recombination, you will end up removing the gene that you wanted to target. However, if the repair process is non homologous end joining (NHEJ), you'll still remove the targeted gene of interest because you'll hopefully have created an indel, rendering the gene product functionally useless.",
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      "title": "",
      "body": "No need to synthesize the plasmid, you can just grow that plasmid in bacteria that have the isotope present in the media. But even then, the results would just say something about plasmid DNA replication :)",
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      "body": "I don't really know much details about biology stuff, so just curious and nievely thought how about:\nSynthesize something like isotopically labelled DNA plasmid (so with smaller number of DNA base pair) would it works? XDD",
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      "permlink": "re-mcw-re-alexanderalexis-the-most-beautiful-experiment-in-biology-20180129t143720898z",
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      "body": "Cool idea, but that would require a lot of money and is not technologically feasible currently, given that synthesized DNA can only be synthesized up to a couple thousand base pairs as limited by the current technology. Additionally, getting such a short DNA molecule to self replicate may prove difficult.\n\nAnother idea is to do polymerase chain reaction (PCR) using isotope labeled dNTPs (the nucleotides A, T, G, and C). Do one round of PCR, save that product, do a second round, save that product, etc until you have enough rounds of PCR (analogous to generations of E. coli) to do your very own Meselson-Stahl inspired experiment!",
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