Detection of integrated murine leukemia viruses in a mouse model of acute myeloid leukemia by fluorescence in situ hybridization combined with tyramide signal amplification.
This study was undertaken to develop a reliable method to enumerate and map somatically acquired, clonal, murine leukemia virus (MuLV) proviral insertions in acute myeloid leukemia (AML) cells from the BXH-2 mouse strain. This was achieved by using fluorescence in situ hybridization combined with tyramide signal amplification (FISH-TSA) and an 8.8 kilobase pair (kb) full-length ecotropic MuLV or 2.0 kb MuLV envelope (env) gene probe. Two-color FISH was utilized combining chromosome-specific probes for regions near the telomere and/or centromere and the MuLV probes. The technique reliably detected germline and somatically acquired, tumor-specific, MuLV proviruses in BXH-2 AML cell lines. It was possible to readily verify homozygous insertions at endogenous ecotropic MuLV loci, Emv1 (chromosome 5), Emv2 (chromosome 8) and a BXH-2 strain-specific locus (chromosome 11). This strategy also verified the presence of molecularly cloned proviral insertions within the mouse Nf1 gene and another locus on distal chromosome 11, as well as on chromosome 7 and chromosome 9 in BXH-2 AML cell line B117. The technique was also used to detect several new tumor-specific, proviral insertions in BXH-2 AML cell lines.
Acar
H
H
University of Minnesota Cancer Center, Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455, USA.
Copeland
N G
NG
Gilbert
D J
DJ
Jenkins
N A
NA
Largaespada
D A
DA
D000208:Acute Disease; D000818:Animals; D002874:Chromosome Mapping / Q000379:methods; D003433:Crosses, Genetic; D015342:DNA Probes; D004251:DNA Transposable Elements / Q000235:genetics*; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D004195:Disease Models, Animal; D005260:Female; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D009052:Leukemia Virus, Murine / Q000235:genetics*; D007951:Leukemia, Myeloid / Q000235:genetics* / Q000821:virology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011533:Proviruses / Q000235:genetics*; D012192:Retroviridae Infections / Q000235:genetics / Q000821:virology; D014412:Tumor Virus Infections / Q000235:genetics / Q000821:virology; D014439:Tyramine; D016662:Virus Integration / Q000235:genetics
D015342:DNA Probes; D004251:DNA Transposable Elements; D004273:DNA, Neoplasm; D014439:Tyramine