Babitsky has been involved in this conflict as a reporter for at least six years. He lived(s) mainly abroad, works for Radio Svoboda and has established a special and trusting relationship with the Chehen "rebels". He has been arrested by the Russian agencies on several ocasions for this type of journalisic activities. For teh Russians, the question was and remains: Is it morally justified to to carry out this line of reporting when you know of all the terror that Basayev brought onto civilians... His (AB's) position is clear - what about Russians' actions in Chechnya... Catch 22 as always...
I have said it over and over again at various discussion about MS Office vs alternatives - for scientists, compatibility between MS Word and Endnote (a dominant citation reference manager) is the single most important reasons to stick with MS. Give me OO, Pages or any other app with built-in capabilities of Endnote, I'd drop MS Office on the spot.
sad, sad news! If you can't change the past or the future why bother travelling in time at all? It is just like going to a museum - see but no touch...
linuxbaby - excellent post! Now, is there any way to know whether any given artist is supported through cdbaby when shopping at iTunes - frankly, I don't mind 99c when most goes to the band but will stick to allofmp3 for all other fat-ass artist ripoffs - alternatively, do you or can you post on your website lists of disributed bands?
Highly efficient endogenous human gene correction using designed zinc-finger nucleases
FYODOR D. URNOV1, JEFFREY C. MILLER1, YA-LI LEE1, CHRISTIAN M. BEAUSEJOUR1, JEREMY M. ROCK1, SHELDON AUGUSTUS1, ANDREW C. JAMIESON1, MATTHEW H. PORTEUS2, PHILIP D. GREGORY1 & MICHAEL C. HOLMES1
1 Sangamo BioSciences, Inc. Pt. Richmond Tech Center 501, Canal Blvd, Suite A100 Richmond, California 94804, USA 2 Department of Pediatrics and Department of Biochemistry, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, Texas 75390, USA
Correspondence should be addressed to M.C.H. (mholmes@sangamo.com) or M.H.P. (matthew.porteus@UTSouthwestern.edu); requests for materials should be addressed to M.C.H.
Permanent modification of the human genome in vivo is impractical owing to the low frequency of homologous recombination in human cells, a fact that hampers biomedical research and progress towards safe and effective gene therapy. Here we report a general solution using two fundamental biological processes: DNA recognition by C2H2 zinc-finger proteins and homology-directed repair of DNA double-strand breaks. Zinc-finger proteins engineered to recognize a unique chromosomal site can be fused to a nuclease domain, and a double-strand break induced by the resulting zinc-finger nuclease can create specific sequence alterations by stimulating homologous recombination between the chromosome and an extrachromosomal DNA donor. We show that zinc-finger nucleases designed against an X-linked severe combined immune deficiency (SCID) mutation in the IL2Rbold italic gamma gene yielded more than 18% gene-modified human cells without selection. Remarkably, about 7% of the cells acquired the desired genetic modification on both X chromosomes, with cell genotype accurately reflected at the messenger RNA and protein levels. We observe comparably high frequencies in human T cells, raising the possibility of strategies based on zinc-finger nucleases for the treatment of disease.
Most human monogenic disorders remain difficult to treat because therapeutic transgenes do not undergo homologous recombination (HR) into the mutated locus1, 2, and gene addition by virus-driven random integration remains a challenge owing to transgene silencing, improper activity or misintegration3, 4. Furthermore, targeted alteration of DNA sequence in vivo--in principle, a powerful basic research technique for studying genome function--in mammals requires sophisticated targeting vectors and drug-based selection1, 2, which limits the use of this approach5-7.
The C2H2 zinc-finger, originally discovered in Xenopus8, is the most common DNA binding motif in all metazoa9. Each finger recognizes 3-4 base pairs of DNA via a single alpha-helix10, 11, and several fingers can be linked in tandem to recognize a broad spectrum of DNA sequences with high specificity12-14. Engineered zinc-finger protein (ZFP)-based DNA binding domains with novel specificities have been extensively applied in vivo to target various effector domains12, 15. Work from the Chandrasegaran laboratory has shown that a ZFP can be coupled to the nonspecific DNA cleavage domain of the Type IIS restriction enzyme, FokI, to produce a zinc-finger nuclease (ZFN)16, which then cuts the DNA sequence determined by the ZFP16, 17. An important specificity mechanism derives from the requirement that two ZFNs bind the same locus, in a precise orientation and spacing relative to each other, to create a double-strand break (DSB; Fig. 1a)17. One mechanism by which eukaryotic cells heal DSBs is homology-directed repair (Fig. 1b)18-20, which transfers information missing at the break from a homologous DNA molecule (Fig. 1b). Work from the Jasin laboratory21, followed by that of others22, 23, demonstrated that the endonuclease I-SceI can potentiate HR into loci previously engineered to contain its own recognition site, and the Carroll24, 25 and Baltimore26 laboratories have shown that a ZFN-invoked DSB increases the rate of HR in model systems.
Note that this observation was made in a laboratory-kept strain of plant. These type of event has little chance in the wild. So, evolution is not an issue here whatsoever. Instead, a more important point here is that such permissive lab conditions allow us to uncover events that are rare or invisible in large or wild life populations. Hopefully this is not an example of missing a related gene in the genome due to incomplete sequencing of very complicated plant genomes (this is issue is mentioned elsewhere in this discussion) but a record of a true fact that has real biological mechanism behind it. Most revolutionary finding in biology since the discovery of DNA as the hereditary substance, namely RNA interference, started with a similar puzzle and also in plants - and it took quite sometime to figure out that it is a general phenomenon throughout the living matter.
This Yahoo decision comes few days after Google's offer of 1 GB to its customers... I think Yahoo simply signes you up for Google account to increase the storage space but uses its crippled interface that gives you an impression that you are in fact using Yahoo... Well done!!!!
90's? How many times we've heard about good then bad effects of coffee? It's same thing now with alcohol - it's a fad no more than that... I say - we live only once so party hard while you have a chance and especially if you are a nerd - there will be only fewer of them chances in the future...
just make sure that your fun does not cause other people's suffering - driving while drunk isn't cool - you wouldn't be writing code while drunk, would you?
I think that Mac Mini $499 defines a new price level at which PC users will now be saying - $499?!?!??! and no flat monitor - this is expensive!!!Macs are still overpriced!
the coolest thing about plos is that every table, figure and any other bit of info has its own URL and can be accessed or linked directly by the reader. In this regard, Plos represents a huge step forward compared to old school of scientific publication.
OK, I admit it - I am crazy about cycling... Now, if you accept that, you'll be able to hear me out - we are as yet to find the best tire out there - the one that will do everything right - corner, hold, being light weight, durable, free from flats, sexy (there I said it). It is being said that cyclists are the trendiest of all - that's true - nowhere else a minimal gain in performance could lead to a win... Tour de France 2003 - over 2300 miles raced - won by seconds... Yes, we need better rubber!!!
Slashdot Mirror
hold back on buying Titanium ... Will wait until Aluminum comes out in couple of years...
First "real and relable" evidence of actual Earth's rotation around the Sun and not vise versa... Kudos Copernicus!
Babitsky has been involved in this conflict as a reporter for at least six years. He lived(s) mainly abroad, works for Radio Svoboda and has established a special and trusting relationship with the Chehen "rebels". He has been arrested by the Russian agencies on several ocasions for this type of journalisic activities. For teh Russians, the question was and remains: Is it morally justified to to carry out this line of reporting when you know of all the terror that Basayev brought onto civilians... His (AB's) position is clear - what about Russians' actions in Chechnya... Catch 22 as always...
the parent is a poorly informed comment and what is so insightful about it escapes me
I have said it over and over again at various discussion about MS Office vs alternatives - for scientists, compatibility between MS Word and Endnote (a dominant citation reference manager) is the single most important reasons to stick with MS. Give me OO, Pages or any other app with built-in capabilities of Endnote, I'd drop MS Office on the spot.
sad, sad news! If you can't change the past or the future why bother travelling in time at all? It is just like going to a museum - see but no touch...
Yes, I did not read TFA
linuxbaby - excellent post! Now, is there any way to know whether any given artist is supported through cdbaby when shopping at iTunes - frankly, I don't mind 99c when most goes to the band but will stick to allofmp3 for all other fat-ass artist ripoffs - alternatively, do you or can you post on your website lists of disributed bands?
Highly efficient endogenous human gene correction using designed zinc-finger nucleases
FYODOR D. URNOV1, JEFFREY C. MILLER1, YA-LI LEE1, CHRISTIAN M. BEAUSEJOUR1, JEREMY M. ROCK1, SHELDON AUGUSTUS1, ANDREW C. JAMIESON1, MATTHEW H. PORTEUS2, PHILIP D. GREGORY1 & MICHAEL C. HOLMES1
1 Sangamo BioSciences, Inc. Pt. Richmond Tech Center 501, Canal Blvd, Suite A100 Richmond, California 94804, USA
2 Department of Pediatrics and Department of Biochemistry, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, Texas 75390, USA
Correspondence should be addressed to M.C.H. (mholmes@sangamo.com) or M.H.P. (matthew.porteus@UTSouthwestern.edu); requests for materials should be addressed to M.C.H.
Permanent modification of the human genome in vivo is impractical owing to the low frequency of homologous recombination in human cells, a fact that hampers biomedical research and progress towards safe and effective gene therapy. Here we report a general solution using two fundamental biological processes: DNA recognition by C2H2 zinc-finger proteins and homology-directed repair of DNA double-strand breaks. Zinc-finger proteins engineered to recognize a unique chromosomal site can be fused to a nuclease domain, and a double-strand break induced by the resulting zinc-finger nuclease can create specific sequence alterations by stimulating homologous recombination between the chromosome and an extrachromosomal DNA donor. We show that zinc-finger nucleases designed against an X-linked severe combined immune deficiency (SCID) mutation in the IL2Rbold italic gamma gene yielded more than 18% gene-modified human cells without selection. Remarkably, about 7% of the cells acquired the desired genetic modification on both X chromosomes, with cell genotype accurately reflected at the messenger RNA and protein levels. We observe comparably high frequencies in human T cells, raising the possibility of strategies based on zinc-finger nucleases for the treatment of disease.
Most human monogenic disorders remain difficult to treat because therapeutic transgenes do not undergo homologous recombination (HR) into the mutated locus1, 2, and gene addition by virus-driven random integration remains a challenge owing to transgene silencing, improper activity or misintegration3, 4. Furthermore, targeted alteration of DNA sequence in vivo--in principle, a powerful basic research technique for studying genome function--in mammals requires sophisticated targeting vectors and drug-based selection1, 2, which limits the use of this approach5-7.
The C2H2 zinc-finger, originally discovered in Xenopus8, is the most common DNA binding motif in all metazoa9. Each finger recognizes 3-4 base pairs of DNA via a single alpha-helix10, 11, and several fingers can be linked in tandem to recognize a broad spectrum of DNA sequences with high specificity12-14. Engineered zinc-finger protein (ZFP)-based DNA binding domains with novel specificities have been extensively applied in vivo to target various effector domains12, 15. Work from the Chandrasegaran laboratory has shown that a ZFP can be coupled to the nonspecific DNA cleavage domain of the Type IIS restriction enzyme, FokI, to produce a zinc-finger nuclease (ZFN)16, which then cuts the DNA sequence determined by the ZFP16, 17. An important specificity mechanism derives from the requirement that two ZFNs bind the same locus, in a precise orientation and spacing relative to each other, to create a double-strand break (DSB; Fig. 1a)17. One mechanism by which eukaryotic cells heal DSBs is homology-directed repair (Fig. 1b)18-20, which transfers information missing at the break from a homologous DNA molecule (Fig. 1b). Work from the Jasin laboratory21, followed by that of others22, 23, demonstrated that the endonuclease I-SceI can potentiate HR into loci previously engineered to contain its own recognition site, and the Carroll24, 25 and Baltimore26 laboratories have shown that a ZFN-invoked DSB increases the rate of HR in model systems.
Figure
Note that this observation was made in a laboratory-kept strain of plant. These type of event has little chance in the wild. So, evolution is not an issue here whatsoever. Instead, a more important point here is that such permissive lab conditions allow us to uncover events that are rare or invisible in large or wild life populations. Hopefully this is not an example of missing a related gene in the genome due to incomplete sequencing of very complicated plant genomes (this is issue is mentioned elsewhere in this discussion) but a record of a true fact that has real biological mechanism behind it. Most revolutionary finding in biology since the discovery of DNA as the hereditary substance, namely RNA interference, started with a similar puzzle and also in plants - and it took quite sometime to figure out that it is a general phenomenon throughout the living matter.
This Yahoo decision comes few days after Google's offer of 1 GB to its customers... I think Yahoo simply signes you up for Google account to increase the storage space but uses its crippled interface that gives you an impression that you are in fact using Yahoo... Well done!!!!
I think they tried to do it to Newton years back...
Maybe that's why it is not a bad idea to have docs with computer skills...
Q: Will bringing down the tracking site stop the flight?
A. I guess we'll see, Mr. Fossett...
Windoze PC off the Net and the problem solved!!!!!
and yes, Apple does some things..just to prove their impracticality...
http://pearpc.sourceforge.net/ - you are welcome to contribute to the project if you are not "completely and utterly inadequate"...
Damn palindrome...
MacOS X... ... but since you are looking for really manly pain of configuring everyting on your own... read the rest of these posts
do the cosmonavts need to show their passports when crossing to the Russian section?
90's? How many times we've heard about good then bad effects of coffee? It's same thing now with alcohol - it's a fad no more than that... I say - we live only once so party hard while you have a chance and especially if you are a nerd - there will be only fewer of them chances in the future... just make sure that your fun does not cause other people's suffering - driving while drunk isn't cool - you wouldn't be writing code while drunk, would you?
Totally dude!
I think that Mac Mini $499 defines a new price level at which PC users will now be saying - $499?!?!??! and no flat monitor - this is expensive!!!Macs are still overpriced!
the coolest thing about plos is that every table, figure and any other bit of info has its own URL and can be accessed or linked directly by the reader. In this regard, Plos represents a huge step forward compared to old school of scientific publication.
cancel his account on iTMS and thus "free" this idiot into the wild
OK, I admit it - I am crazy about cycling... Now, if you accept that, you'll be able to hear me out - we are as yet to find the best tire out there - the one that will do everything right - corner, hold, being light weight, durable, free from flats, sexy (there I said it). It is being said that cyclists are the trendiest of all - that's true - nowhere else a minimal gain in performance could lead to a win... Tour de France 2003 - over 2300 miles raced - won by seconds... Yes, we need better rubber!!!