First synthetic virus created

BBC ^ | July 11, 2000 | Dr David Whitehouse

[Nebullis]

Scientists have assembled the first synthetic virus.

The US researchers built the infectious agent from scratch using the genome sequence for polio.

Read More...

[Pyro7480]

What convinces you that my judgement is based SOLELY on my faith? I have 4 leftist friends of mine who have "alternative" beliefs or even atheistic, but they have come to the same conclusion concerning this and other similar scientific endeavours. This is because we have all thought over this carefully, and have come to this conclusion. Science fiction writers (while I don't cite them as a source of morality) have written stories about similiar scenarios, and since humans are IMPERFECT, we can potentially destroy all that we have endeavoured in creating up to this point if this technology is pursued any further.

[Ronin]

Yeah, I got that giggle too. I said much the same thing when the story first came out.

Cloning rabbits would be funnier -- but not by much.

[gcruse]

You're right. I'm sorry.  I shot from the hip.  I was strongly
reminded of an argument over whether inoculating someone
against, say, smallpox was playing god.  It supposedly
thwarted some heavenly dictate that we would die from
the disease when we were supposed to.  The very act
of giving someone a mild form of the disease even
though it protected them from it in the long run was
also akin to destroying the technology in question
even though much good could come from it because
it involves doing something counter to conventional
thinking in the early stages of research.

[Pyro7480]

I think that's not comparable to the recent development. Jenner wasn't trying to create a virus, he was trying to prevent one from killing people by using cowpox.

[T. P. Pole]

Let me see if I understand this. They use a recipe found on the internet, then claim that they were the first to do it?

If that's the case, can I claim that I made the first batch of chocolate chip cookies because I used a recipe found on the bag of chips?

[Libertarianize the GOP]

.

[gcruse]

I think that's not comparable to the recent development. Jenner
wasn't trying to create a virus, he was trying to prevent one from
killing people by using cowpox.

He gave people the disease in order to
prevent them from getting it later. His first
volunteer was at least as brave as the first
person to ever eat an oyster.

Or similarly, the guys who developed the
nuclear bombs that ended WWII weren't
sure that the first test detonation chain
reaction wouldn't consume the entire atmosphere.

In both these cases, good came out of
spooky technology.
 
 

[neutrino]

The next step will be to combine traits of several viruses. Perhaps a fragment as contagious as the flu - combined with something deadly. Maybe a little fragment that produces a lethal toxin.

Sleep well....

[Pyro7480]

This is very true. Perhaps I was also a bit too extreme when I said the technology should be destroyed, BUT this technology should be very closely monitored before evil forces get a hold of it.

[William Tell]

neutrino said: "Perhaps a fragment as contagious as the flu - combined with something deadly. Maybe a little fragment that produces a lethal toxin. "

Or similarly, a fragment as contgagious as the flu - but incapable of causing serious illness, and combined with genes which give the "infected" permanent protection against the one hundred most common types of cancer.

[aruanan]

Dr Wimmer said assembling the polio virus showed that eradicating a virus in the wild might not mean it was gone forever because biochemists could now reconstruct those viruses from blueprints.

You could build the smallpox virus genome from scratch and then put it into another viral capsid (1), one that would be easily internalized, such as an adenovirus (2), as used for gene therapy and one of the five major virus families that cause the common cold; once internalized by the host, perhaps as easily as being picked up from a doorknob like a cold is at the office during cold and flu season, the ensuing viral replication would then result in the production of a lot of native smallpox in the native smallpox capsid and convert into a normal mode of transmission. Such an agent could be spread intially throughout large office buildings by cleaning personnel at night. Over the course of a week or so, before people caught on to the fact that the disease was smallpox and got an inkling of its initial mode of infection, large numbers of public places (armrests on airside chairs at airports, computer keyboards in public schools and office buildings, desktops, even kindergarten crayons, etc.) could be contaminated, and many first generation cases started. If the initial infection is widespread, sudden, and unanticipated, the resulting numbers of secondary cases could be enough to cause major problems. Various terrorist scenarios have been proposed for such modes of transmission as 3 aerosol bombs infecting 1000 people in each of 3 shopping malls or 5 al Qaeda terrorists (Science Magazine: BIOTERRORISM: How Devastating Would a Smallpox Attack Really Be? by Martin Enserink). The authors conclude that the resulting infection would be small. But a recombinant smallpox genome in an adenovirus or rhinovirus capsid spread quietly could be a different story entirely. Imagine, instead of 5 al Qaeda terrorists, 100 janitors at large junior and senior high schools and another 20 or 30 housekeeping staff at major airports.

1.

J Virol 1998 Dec;72(12):10298-300
In vitro construction of pseudovirions of human papillomavirus type 16: incorporation of plasmid DNA into reassembled L1/L2 capsids.

Kawana K, Yoshikawa H, Taketani Y, Yoshiike K, Kanda T.

Division of Molecular Genetics, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan.

Lack of permissive and productive cell cultures for the human papillomaviruses (HPVs) has hindered the study of virus-neutralizing antibodies and infection. We developed a cell-free system generating infectious HPV16 pseudovirions. HPV16 L1/L2 capsids, which had been self-assembled in insect cells (Sf9) expressing virion proteins L1 and L2, were disassembled with 2-mercaptoethanol (2-ME), a reducing agent, and reassembled by removal of 2-ME in the presence of a beta-galactosidase expression plasmid. Plasmid DNA purified together with the reassembled capsids was resistant to DNase I digestion. The reassembled pseudovirions mediated DNA transfer to COS-1 cells, as monitored by induced beta-galactosidase activity. Transfer was inhibited by anti-HPV16 L1 antiserum but not by antisera against L1s of HPV6 and HPV18. Construction in vitro of HPV pseudovirions containing marker plasmids would be potentially useful in developing methods to assay virus-neutralizing antibodies and to transfer exogenous genes to HPV-susceptible cells.

PMID: 9811779 [PubMed - indexed for MEDLINE]

J Gen Virol 1999 Jan;80 ( Pt 1):39-46
The major capsid protein, VP1, of human JC virus expressed in Escherichia coli is able to self-assemble into a capsid-like particle and deliver exogenous DNA into human kidney cells.

Ou WC, Wang M, Fung CY, Tsai RT, Chao PC, Hseu TH, Chang D.

Department of Microbiology, Chung Shan Medical and Dental College, Taichung, Taiwan, Republic of China.

The full-length major capsid protein, VP1, of the human polyomavirus JC virus was cloned and expressed in Escherichia coli. VP1 protein expressed in E. coli self-assembled into capsid-like particles and caused haemagglutination of human O-type red blood cells. Caesium chloride density-gradient centrifugation analysis revealed that the capsid-like particles consisted of virion-like pseudovirion and empty capsid-like pseudocapsid populations. The morphology of pseudo-virion and pseudocapsid particles was observed under the electron microscope. The pseudovirions contained DNA and RNA molecules but the pseudocapsids did not contain any nucleic acid, as analysed by DNA extraction. DNA-binding activity of VP1 was also demonstrated by the South-Western probing method in vitro. Furthermore, pseudocapsids were able to deliver exogenous DNA into human foetal kidney epithelial cells. These results indicate that recombinant JC virus VP1 is able to self-assemble into capsid-like particles and to package DNA in the absence of the minor capsid proteins, VP2 and VP3. This prokaryotic assembly system may facilitate the investigation of maturation mechanism(s) of polyomaviruses. Furthermore, capsid-like particles of JC virus VP1 generated in E. coli potentially could be used as a human gene transfer vector.

PMID: 9934681 [PubMed - indexed for MEDLINE]

2.

Adenoviruses develop within the nuclei of infected cells, where they often can be observed packed in an apparently crystalline arrangement. In humans, adenoviruses cause acute mucous-membrane infections of the upper respiratory tract, the eyes, and frequently the regional lymph nodes, bearing considerable resemblance to the common cold. Like the cold viruses, adenoviruses are often found in latent infections in clinically healthy persons. There are more than 45 different members of the adenovirus group, but only a few commonly cause illness in humans; it is thus possible to prepare a vaccine against these viruses. In contrast, there are more than 100 cold viruses, all of which are fairly commonly found as disease agents; this great number makes the development of a vaccine for the common cold virtually impossible.

J Neurosci Methods 2002 Feb 15;114(1):99-106
High-efficiency adenovirus-mediated in vivo gene transfer into neonatal and adult rodent skeletal muscle.

Sapru MK, McCormick KM, Thimmapaya B.

Molecular Neuroscience Laboratory, Departments of Kinesiology and Psychiatry, University of Illinois at Chicago, MC 194, 901 West Roosevelt Road, Chicago, IL 60608, USA. msapru@uic.edu

Several methodological limitations have emerged in the use of viral gene transfer into skeletal muscle. First, because the nuclei of mature muscle fibers do not undergo division, the use of strategies involving replicative integration of exogenous DNA is greatly limited. Another important limitation concerns the maturation-dependent loss in muscle fiber infectivity with adenoviral vectors. In this study, we investigated the possibility that high-titer infections with recombinant adenovirus, expressing a foreign marker gene under the control of a strong viral promoter, can significantly improve the efficiency of gene transfer in vivo into neonatal and adult rat skeletal muscle. High-titer (2 x 10(10) plaque forming units) intramuscular injection of replication-defective adenovirus vector, expressing green fluorescent protein (GFP) under the control of cytomegalovirus promoter, resulted in GFP expression in 99 +/- 0.34% of fibers in the adult soleus muscle and in approximately 85 +/- 1.44% of fibers in the adult tibialis anterior muscle. Interestingly, reduction in injected adenoviral dose significantly reduced the number of GFP-positive fibers in the adult tibialis anterior muscle, but not in the soleus muscle. However, in neonates, adenoviral infection resulted in GFP expression in 96-99% of the fibers in the tibialis anterior and the gastrocnemius muscles regardless of administered adenoviral dose.

PMID: 11850044 [PubMed - indexed for MEDLINE]

[Nebullis]

Viral delivery systems are commonly used for delivery of specific genes or even proteins. Packaging a large, enveloped virus, in a small capsid, with hopes of viability or pathogenicity is not of concern at this time.

[Nebullis]

Thanks, sourcery. I searched some obvious key words, but the title isn't very forthcoming.

[Barnacle]

"The reason we did it was to prove that it can be done and it now is a reality," said Dr Eckard Wimmer

And like the four-minute mile, no one could do it until someone did it, then everyone could do it.

This is bad Voodoo for sure.

[RightWhale]

When they say "from scratch", what is "scratch"? Amino acids?

[Equality 7-2521]

The next step will be to combine traits of several viruses. Perhaps a fragment as contagious as the flu - combined with something deadly. Maybe a little fragment that produces a lethal toxin.

Yeah, and maybe the survivors will start to have dreams about an old granny lady from Hemingford Home, Nebraska. And then they'll all either move to Boulder or Las Vegas. Then maybe some deranged pyromaniac will find a nuclear weapon out in the desert and bring it back to Vegas and the hand of God will ignite it.

[aruanan]

Packaging a large, enveloped virus, in a small capsid, with hopes of viability or pathogenicity is not of concern at this time.

The question wasn't of large versus small but of the concept of using one viral capsid to deliver the artificially-constructed genome of another virus. Although that may not be a concern at "this time", it should be a concern for a time not too far off.

[Nebullis]

When they say "from scratch", what is "scratch"? Amino acids?

And nucleic acids. I don't have any more information about this particular experiment, but, in general, coat proteins can be synthesized and in the right solution will spontaneously form the right capsid shape. RNA (and DNA) sequences are synthesized in the lab on automated synthesizers.

[aruanan]

When they say "from scratch", what is "scratch"? Amino acids?

No, you'd start with constructing the genome from nucleic acids via PCR.

[LarryLied]

Did they select polio because it is the easiest virus to make or because their work will receive more publicity?