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Reading the methods section, it looks the T-cells infected with HIV-1 that are used for targeted for gene editing came from a cell line and incubated in a cultu
by noname123 11y ago
Reading the methods section, it looks the T-cells infected with HIV-1 that are used for targeted for gene editing came from a cell line and incubated in a cultured medium.
For people who are involved in dealing with CRISPR/Cas9, what is the effective rate of gene editing done in vitro vs. in vivo?
Would the molecular machinery of CRISPR/Cas9 encounter any natural defense mechanism of the human immune system when attempting to mutate target sites on T-cells in say, lymph nodes of adult humans (contrasting to changing DNA in the germ-lines of bacteria or even human embryo's)?
- noipv4 11y agoThis link might be useful for answering your question: http://www.nature.com/nature/journal/v514/n7522/abs/nature13589.html http://www.nature.com/nature/journal/v514/n7522/abs/nature13...
- cryoshon 11y ago>For people who are involved in dealing with CRISPR/Cas9, what is the effective rate of gene editing done in vitro vs. in vivo? Difficult question. CRISPR is ironclad in vitro, but in vivo it's not well understood. For reference, other methods of gene transfer/editing are almost exclusively for in vitro use, with the exception of lentiviral vector-based transduction. >Would the molecular machinery of CRISPR/Cas9 encounter any natural defense mechanism of the human immune system when attempting to mutate target sites on T-cells in say, lymph nodes of adult humans (contrasting to changing DNA in the germ-lines of bacteria or even human embryo's)? Yes. The cell membrane, the nuclear envelope within the cell, and the chromatin/chromosomal structure of the genetic material within the nuclear envelope are huge barriers that can't be skipped across easily when in vivo because many of the convenient work-arounds aren't suitable for in vivo use. Bacteria are always much easier.
- L_Rahman 11y agoThe study used Lentivirus as the delivery mechanism, but ex vivo. I imagine an in vivo study would be the next step, and I'm interested in seeing its outcome.
- carbocation 11y ago> For people who are involved in dealing with CRISPR/Cas9, what is the effective rate of gene editing done in vitro vs. in vivo? It will depend in part on the tissue of interest and the delivery vector. In the mouse liver, the Broad group under Zhang has gotten efficiencies as high as 40% using Staphylococcal Cas9 [1]. 1 = http://www.nature.com.sci-hub.io/nature/journal/v520/n7546/full/nature14299.html http://www.nature.com.sci-hub.io/nature/journal/v520/n7546/f...
- itcmcgrath 11y agoI found this + the op's comments fascinating purely because I wouldn't have been able to tell if both were real, or just fake technical talk of the ilk my friends and I sometimes do with meaningless software/hardware jargon as a joke, without Googling.
- noname123 11y agoWell, for the record, I am a programmer working at the Broad, a genomics center that is mired in the current CRISPR patent dispute (although I don't work with CRISPR at all). The answerer I believe is a MD/PhD at Mass General Hospital and before that a research scientist at the Broad working on genomics (and so is the author of the cited paper on mouse liver, small world here on HN). So I can be full of BS, but I don't think the answerer is. Tbh, I intentionally added a few more jargons more than necessary (I could have just asked "Can gene editing be effective in live humans T-cells vs. cells on a petri dish?") to field my question; but I intentionally did so to filter out the pop-sci, speculative futurists to get answers from people in life sciences research.