Mutagenesis, insertional

There are two strategies to use insertional mutagenesis for cloning purposes. One is the direct tagging approach. In this case a mutant, for example a spontaneous or chemically induced mutant, has already been identified, but no information on the nature of the mutated gene is available. 

The mutant is then crossed with a wild-type line carrying an active transposable element, and the F] progeny is screened for the presence of a plant with the mutant phenotype. Such a mutant will only be identified if the wild-type allele of the gene of interest has been mutated, presumably as the result of an insertion. 

---

The development of leukemia in patients receiving retrovirally transduced cells, clearly underlines that insertional mutagenesis is a major concern for integrating vectors. 

For intracellular immunization strategies of hepatitis B and C, lentiviral vectors have been tested (Matskevich et al. 2003 Parouchev et al. 2006). However, as the turnover of hepatocytes is thought to be much slower than that of HPC or T cells, nonintegrating vectors that carry a reduced risk of insertional mutagenesis are favored (Nowak et al. 1996). 

Baltimore D (1988) Gene therapy. Intracellular immunization. Nature 335 395-396 Basta S, Stoessel R, Easier M, van den Broek M, Groettrup M (2005) Cross-presentation of the long-lived lymphocytic choriomeningitis virus nucleoprotein does not require neosynthesis and is enhanced via heat shock proteins. J Immunol 175 796-805 Baum C (2007) Insertional mutagenesis in gene therapy and stem cell biology. Curr Opin Hematol 14 337-342... 

AZPIROZ, R., FELDMANN, K.A., T-DNA insertion mutagenesis in Arabidopsis Going back and forth, 1997, Trends Genet., 13 152-156. 

Determination of potential for insertional mutagenesis (malignant transformation). 

Boison, G., Schmitz, O., Mikheeva, L., Shestakov, S. and Bothe, H. (1996) Cloning, molecular analysis and insertional mutagenesis of the bidirectional hydrogenase genes from the cyanobacterium Anacystis nidulans. FEES Lett., 394, 153-8. 

Retroviral insertional mutagenesis has been used for more than 2 decades to identify proto-oncogenes. More recently, a causal role of retroviral... 

Modlich U., Kustikova O.S., Schmidt M., Rudolph C., Meyer J., Li Z., Kamino K., von Neuhoff N., Schlegelberger B., Kuehlcke K., Bunting K.D., Schmidt S., Deichmann A., von Kalle C., Fehse B., Banm C. (2005) Lenkemias following retroviral transfer of multidrug resistance 1 (MDRl) are driven by combinatorial insertional mutagenesis. Blood Mi, 4235-4246. 

Dn Y., Spence S.E., Jenkins N.A. and Copeland N.G. (2005) Cooperating cancer-gene identification throngh oncogenic-retrovirns-indnced insertional mutagenesis. Blood 106, 2498-2505. 

---