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I find it interesting that although anyone can experiment with CRISPR in their living room using something like this http://www.the-odin.com/ that no one has just tried modifying the non-coding DNA a lot and then observed any changes (or lack thereof) in the specimen.
Earlier quoted context omitted.
> "Co-housed FVB/NJ mice without CRISPR-mediated correction were used as the functional-deficient control. Briefly, an sgRNAexpressing plasmid had been coinjected, into FVB/NJ zygotes, with the single-stranded oligodeoxynucleotide (ssODN) donor template and Cas9 protein to generate mosaic F0 founders.1" Following to reference 1: > "The sgRNA plasmid was co-injected with the single-stranded oligodeoxynucleotide (ssODN…
The more you work in the field, the more you discover how underwhelming the details in biology papers are. They give results without detailing the algorithms, hence destroying reproducibility, they hide datasets behind confidentiality, presents large-scale graphs without precise data, and so on. Generally, reproducibility in biology papers is but a far away dream.
Pre-publication press release. Full text of article not available. How good was their control group? The press release says they used mice that had previously been edited with CRISPR/Cas9 and found off-target mutations, most of which were single nucleotides. Did they have a good control group so as to exclude random point mutations? Who knows? What is the theory on how CRISPR/Cas9 causes single nucleotide off-target…
I'm just a normal person with nothing to do with this, and I expected such "surprises" from the beginning.
I remain convinced mankind should not mess with this.
Most of the DNA that suffers random mutations is non-coding DNA. It's completely unknown what effects, if any, modifying non-coding DNA might have as its use is unknown. I find it interesting that although anyone can experiment with CRISPR in their living room using something like this http://www.the-odin.com/ that no one has just tried modifying the non-coding DNA a lot and then observed any changes (or lack thereof…
You need probably low 4-figures of equipment first; a -20C freezer for DNA/buffer storage, small centrifuge, thermal cycler, some way of getting genes into a target, and incubation space for whatever you're modifying. Preferably you'll also have consistent temperature control and an extremely clean environment. You also need a source of primer sequences, which are short customized sequences of DNA used to tl;dr get a lot of copies of a DNA sequence which you only have in small quantities.
Nothing too difficult, and much of that equipment can be DIY'd for the home lab. But many chemical suppliers also won't ship to residential addresses or onboard individuals as customers, so you'll need to incorporate and shop around a bit. And wet lab protocols can be very finicky; you'll probably need to run through your workflow a lot before you get to the point where you can semi-reliably go from start to finish without fucking up. Often, you won't know exactly how you fucked up, but you can't argue with a lack of results.
It's possible, I think, but it's also time-consuming and difficult.
Anyways, if you did get a reliable setup working with say, micropropagated plant specimens, there are far more interesting prizes than seeing what non-coding DNA does. Plants make all kinds of cool stuff, from scents to flavors to alkaloids. And they've demonstrated an ability to take genes from things like jellyfish for e.g. autoluminescence, too.
I seem to recall that in prior HN stories about CRISPR some poster was saying he believed CRISPR didn't work the way people thought it worked. He said it was simply killing cells that didn't have the desired mutation and biologists weren't realising that because of design errors in the experiments (or rather, sometimes mass cell dieoffs were being reported but not dwelled upon). If CRISPR isn't actually editing the D…
Don't think so. Originally the CRISPR mechanism was discovered in bacteria, which only have one cell. It doesn't make sense for bacteria's anti-virus system to kill it's only cell.
After all a colony of bacteria is (usually) a set of identical clones. As long as a few survive, the DNA lives on.
Pre-publication press release. Full text of article not available. How good was their control group? The press release says they used mice that had previously been edited with CRISPR/Cas9 and found off-target mutations, most of which were single nucleotides. Did they have a good control group so as to exclude random point mutations? Who knows? What is the theory on how CRISPR/Cas9 causes single nucleotide off-target…
Most of the DNA that suffers random mutations is non-coding DNA. It's completely unknown what effects, if any, modifying non-coding DNA might have as its use is unknown. I find it interesting that although anyone can experiment with CRISPR in their living room using something like this http://www.the-odin.com/ that no one has just tried modifying the non-coding DNA a lot and then observed any changes (or lack thereof…
It's still fairly hard to do in a living room, if you want to do anything novel. You need probably low 4-figures of equipment first; a -20C freezer for DNA/buffer storage, small centrifuge, thermal cycler, some way of getting genes into a target, and incubation space for whatever you're modifying. Preferably you'll also have consistent temperature control and an extremely clean environment. You also need a source of…