Measurement of SIL-TAL1 fusion gene transcripts associated with human T-cell lymphocytic leukemia by real-time reverse transcriptase-PCR.
TAL1 disruption at 1p32 [del(1p)] is a common rearrangement in the development of T-cell acute lymphocytic leukemia (T-ALL). The del(1p) are usually interstitial 90kb deletions placing TAL1 under control of the SCL interrupting locus (SIL) gene forming the SIL-TAL1 fusion product. A reverse transcriptase real-time PCR assay to quantify SIL-TAL1 fusion genes is described. A SIL-TAL1 fusion gene RNA transcript was built that permitted absolute standard curves to be generated. Sensitivity of the RT-PCR assay was determined to be 10 cells (CEM cell line) in 10(6) human lymphocytes. Peripheral blood lymphocytes from 10 healthy adults and 10 neonates were assayed. None of the samples showed any SIL-TAL1 expression. However, when lymphocytes from three adults were cultured in vitro the SIL-TAL1 transcript was detectable in the RNA isolates. No RAG2 expression was detected in these expanded samples, suggesting that the clones bearing the SIL-TAL1 fusion gene may have existed at low levels prior to the ex vivo expansion.
Curry
John D
JD
Division of Immunology, Department of Molecular and Cellular Biology, University of California at Berkley, 439 Life Sciences Addition, Berkeley, CA 94720-3200, USA.
Smith
Martyn T
MT
D000328:Adult; D051792:Basic Helix-Loop-Helix Proteins; D002478:Cells, Cultured; D019457:Chromosome Breakage; D002878:Chromosomes, Human, Pair 1 / Q000235:genetics; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D018628:Gene Dosage; D018257:Helix-Loop-Helix Motifs; D006801:Humans; D007231:Infant, Newborn; D047908:Intracellular Signaling Peptides and Proteins; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000235:genetics*; D008214:Lymphocytes; D009687:Nuclear Proteins; D015514:Oncogene Proteins, Fusion / Q000235:genetics; D010957:Plasmids; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D000073942:T-Cell Acute Lymphocytic Leukemia Protein 1; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D045567:VDJ Recombinases
D051792:Basic Helix-Loop-Helix Proteins; D004254:DNA Nucleotidyltransferases; D017931:DNA Primers; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D015514:Oncogene Proteins, Fusion; D011506:Proteins; D011518:Proto-Oncogene Proteins; D011993:Recombinant Fusion Proteins; D000073942:T-Cell Acute Lymphocytic Leukemia Protein 1; D014157:Transcription Factors; D045567:VDJ Recombinases; C499311:RAG2 protein, human; C071236:STIL protein, human; C064655:V(D)J recombination activating protein 2; C067143:TAL1 protein, human