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New molecular tools help terminate HIV in cultured human cells

Scientists have devised a way to delete the HIV-1 proviral DNA from cultured human cells, offering a potentially more permanent method of ridding the HIV virus.
 
Temple University scientists created "molecular tools" that can hunt the viral genome and terminate the HIV-1 DNA. 
 
The team -- led by Kamel Khalili, PhD, Professor and Chair of the Department of Neuroscience at Temple University -- targeted the HIV virus by combining a DNA-snipping enzyme called a nuclease and a targeting strand of RNA called a guide RNA (gRNA).
 
The team created a 20-nucleotide strand of gRNA to target the HIV-1 DNA. The strands were next injected into HIV-infected cells, where they were able to extract the 9,709-nucleotides that make up the HIV-1 genome. The cell's gene repair machinery then fastens the loose ends of the genome back together, leaving it HIV free.
 
What's more is that since the gRNA strands don’t contain human DNA sequences, the host cells are protected from further cellular DNA damage.


The team found that this particular method was effective in several cell types that can harbor HIV-1, such as microglia and macrophages, as well as in T-lymphocytes.
 
"We are working on a number of strategies so we can take the construct into preclinical studies," said Khalili. "We want to eradicate every single copy of HIV-1 from the patient. That will cure AIDS. I think this technology is the way we can do it."
 
This particular method isn't quite ready for the clinic yet because the team must find a method of delivering the therapeutic agent to every single infected cell. Also, because HIV-1 can mutate, treatment may need to be individualized for each patient's viral sequences.
 
However, the research looks to be a potential upgrade from current methods. As of now, patients have to take a lifelong drug regimen to control the virus and prevent new attacks.
 
HIV research in the past has also proved to be promising, but still hasn’t hit the mainstream. For instance, doctors at the Dana-Farber/Brigham and Women's Cancer Centre in Boston performed bone marrow transplants on two men with HIV, with neither needing to use AIDS drugs for extended periods of time since the operations. 
 
In 2013, researchers from Oregon Health and Science University (OHSU) found that a vaccine cleared an aggressive form of HIV in half of all monkeys treated. 

Source: Temple University



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CRISPR tech
By sgw2n5 on 7/22/2014 2:09:06 PM , Rating: 2
Within the last year, CRISPR technology has really taken off. It is trivially easy to edit a genome now, but the catch is you have to edit the genome at the 1-cell stage (through microinjection of the CRISPRs), or transfect the CRISPR into cultured cells.

The hard part will be finding a way transfect living tissue (or lymphocytes) with the CRISPRs, and ensuring that you get them all.




RE: CRISPR tech
By daboom06 on 7/22/2014 2:19:18 PM , Rating: 2
you literally reworded a sentence from the article. good job.

the actual comment here is: it's hilarious to say a problem's solved all except for the last step, which happens to also be a decades-old impossibility.

seriously, getting rna to every cell in the body? to every cell in an organ? there's rnase everywhere just waiting to rip it all to shreds AND intercellular structures can completely isolate huge numbers of cells.


RE: CRISPR tech
By sgw2n5 on 7/22/2014 2:53:24 PM , Rating: 2
quote:
you literally reworded a sentence from the article. good job.

Really? Which one?

quote:
seriously, getting rna to every cell in the body? to every cell in an organ? there's rnase everywhere just waiting to rip it all to shreds AND intercellular structures can completely isolate huge numbers of cells.

Why would you want to get this particular gRNA+Cas9 RNA into each cell? This is HIV... the viral genome will not be in every cell, just lymphocytes. This is much more realistic of a target than knocking out a some hypermorphic allele that causes a full blown genetic disorder.

But yeah... there have been several genome editing technologies that work very well (TALENs were all the rage just 3 years ago, and ZFNs a few years before that), but delivery in living tissue has ALWAYS been the barrier.


RE: CRISPR tech
By Reclaimer77 on 7/22/2014 4:51:04 PM , Rating: 2
I have no idea what the fuck you guys are talking about.

Just get to the part where I can fuck random ppl without these damn rubbers, plx.


RE: CRISPR tech
By foxalopex on 7/23/2014 3:40:35 PM , Rating: 2
The answer would be no. Even if you could get perfect treatment for AIDS, there's still other diseases to worry about besides AIDS that could seriously mess up your life. It's possible to get cancer via HPV for example. Not to mention what to do in an unplanned pregnancy.


RE: CRISPR tech
By kamk44 on 7/23/2014 11:37:36 PM , Rating: 2
Simple, just create one of those Star Trek transporters and then when you get beamed somewhere and reassembled leave out the bad parts like cancer. Have some extra material on hand to fill it in with healthy cells (and maybe a bigger penis or breasts...oh the possibilities).


Elysium
By havoti97 on 7/22/2014 11:16:09 PM , Rating: 2
Sadly, potential life saving techs like this will likely only be catered to the Bill Gates and Steve Jobs of this world... a la Magic Johnson. I don't think this will be available for everyone in the US, much less the world, for some time to come.




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