Targeted mutagenesis

Faruqi a.F., Egholm M., Glazer P.M. Peptide nucleic acid-targeted mutagenesis of a chromosomal gene in mouse cells. Proc. Natl Acad. Sci. USA 1998 95 1398-1403. 

Several other fluorescent proteins have been isolated from a variety of species. We will briefly discuss the variants with the most optimal spectroscopic properties. A true orange fluorescent protein was isolated from the stony coral Fungia concinna, with good absorbance and a quantum yield of 0.45. Targeted mutagenesis was used to monomerize the protein, yielding mKO with an absorbance of 51,600 M-1 cm 1 and a quantum yield of 0.74 [84, 85], The crystal... 

Rajagopalan, L.E., Burkholder, J.K., Turner, J., Culp, J., Yang, N.-S. and Malter, J.S. (1995) Targeted mutagenesis of GM-CSF cDNA increases transgenic mRNA stability and protein expression in normal cells. Blood, 86, 2551-2558. 

Spicer, A.P. et al., Investigation of hyaluronan function in the mouse through targeted mutagenesis, Glycoconj. J., 19, 341, 2002. 

Simpson EM, Linder CC, Sargent EE, Davisson MT, Mobraaten LE, Sharp II (1997) Genetic variation among 129 substrains and its importance for targeted mutagenesis in mice. Nat Genet 76 19-27. 

Such long-distance mutations typically do not strongly influence the substrate acceptance profile of enzymes. In fact, it appears that, for effectively altering the substrate specificity of an enzyme, one should preferentially target active site residues. Such a targeted mutagenesis of first shell residues has been shown to result in dramatic changes in substrate specificity and/or enantioselectivity [49]. However,... 

Fleming RE, Ahmann JR, Migas MC, Waheed A, Koeffler HP, Kawabata H, et al. Targeted mutagenesis of the murine transferrin receptor-2 gene produces hemochromatosis. Proc Natl Acad Sci USA 2002 99 10653-8,... 

Waterkeyn, J. G., Wickham, M. E., Davern, K. M., Cooke, B. M., Coppel, R. L., Reeder, J. C., Culvenor, J. G., Waller, R. F., and Cowman, A. F. (2000). Targeted mutagenesis of Plasmodium falciparum erythrocyte membrane protein 3 (PfEMP3) disrupts cytoadherence of malaria-infected red blood cells. EMBO ]. 19, 2813-2823. 

The disadvantages of these methods are similar to all inhibitors and activators however the effect is quite likely only partial, and there may be unwanted side effects on other molecules. Consequently, in the future genetic alteration by targeted mutagenesis will be an indispensable tool for both in vivo and in vitro approaches to bio-pharmaceutical research. 

Wood, R. D., and Hutchinson, F. (1984). Non-targeted mutagenesis of unirradiated lambda phage in Escherichia coli host cells irradiated with ultraviolet light. J. Mol. Biol. 173, 293-305. 

Cooksley, C.M., Zhang, Y, Wang, H., Redl, S. et al (2012) Targeted mutagenesis of the Clostridium acetobutylicum acetone-butanol-ethanol fermentation pathway. Metab. Eng., 14, 630-641. 

Computational analysis Targeted mutagenesis/funi ional assay f 1 Epitope 1 identification... 

Nagy, A. and Rossant, J. (1996) Targeted mutagenesis Analysis of phenotype without germ line transmission. J. Clin. Invest. 97,1360-1365. 

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