15 November 2007

Transgenic methods

One of the exciting prospects of Marmorkrebs is that any genetic change to an individual should be propagated to all its offspring, so changes introduced by mutations and transgenics would be easily maintained.

Vogt and colleagues (2004) note that Sarmasik and collegues (2001) had successfully transformed the crayfish species Procambarus clarkii. Sarmasik and colleagues based their methods on those of Burns and colleagues (1993) and Yee and colleagues (1994). All of which tells you how to design this stuff from scratch.

Fortunately, this does not appear to be necessary. A pantropic retroviral expression system is available commercially from ClonTech, which seems to be based on the work of Burns and colleagues.

Note, however, that because this is a viral method of delivery, it has a higher biosafety level than many labs might have.

References

Burns JC, Friedmann T, Driever W, Burrascano M, Yee J. 1993. Vesicular stomatitis Virus G glycoprotein pseudotyped retroviral vectors: Concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells. Proceedings of the National Academy of Sciences 90(17): 8033-8037.
http://www.pnas.org/cgi/content/abstract/90/17/8033

Sarmasik A, Chun CZ, Jang I-K, Lu JK, Chen TT. 2001. Production of transgenic live-bearing fish and crustaceans with replication-defective pantropic retroviral vectors. Marine Biotechnology 3: S177-S184.
http://dx.doi.org/10.1007/s10126-001-0040-3

Sarmasik A, Jang I-K, Chun CZ, Lu JK, Chen TT. 2001. Transgenic live-bearing fish and crustaceans produced by transforming immature gonads with replication-defective pantropic retroviral vectors. Marine Biotechnology 3(5): 470-477.
http://dx.doi.org/10.1007/s10126001-0019-0

Yee J, Miyanohara A, LaPorte P, Bouic K, Burns JC, Friedmann T. 1994. A General method for the generation of high-titer, pantropic retroviral vectors: Highly efficient infection of primary hepatocytes. Proceedings of the National Academy of Sciences 91(20): 9564-9568.
http://dx.doi.org/10.1073/pnas.91.20.9564

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