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Biological microchip for establishing the structure of fusion transcripts involving MLL in children with acute leukemia
MLL is involved in fusion genes with more than 100 partner genes, approximately 80 of which have been characterized at the molecular level. MLL fusion genes are often found in infants (60–80% of acute lymphoblastic leukemia (ALL) cases and 40–50% of acute myeloblastic leukemia (AML) cases) and are a...
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Published in: | Molecular biology (New York) 2016-11, Vol.50 (6), p.852-859 |
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Main Authors: | , , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | MLL
is involved in fusion genes with more than 100 partner genes, approximately 80 of which have been characterized at the molecular level.
MLL
fusion genes are often found in infants (60–80% of acute lymphoblastic leukemia (ALL) cases and 40–50% of acute myeloblastic leukemia (AML) cases) and are appreciably rarer (8–10%) in children older than 1 year of age.
MLL
rearrangements are important markers in diagnosis and treatment choice. To identify the partner gene is of primary importance for prognosis and minimal residual disease monitoring. The structure of the fusion gene, including localization of the
MLL
breakpoints, is also informative. A method was developed to examine the fusion transcripts in order to identify the partner gene among the six most common ones and to establish the exon structure of the rearranged
MLL
. The method includes a multiplex reverse transcriptase–polymerase chain reaction (RT–PCR) to amplify and to fluorescently label a fusion transcript fragment and subsequent hybridization of the product on a biological microchip with immobilized oligonucleotides complementary to exons of
MLL
and its partner genes
AFF1
,
MLLT1
,
MLLT3
,
MLLT4
,
MLLT10
, and
ELL
. Hybridization results were verified by sequencing the RT–PCR products and, in some cases, performing long-distance inverse PCR (LDI-PCR). The study involved 38 bone marrow samples from ALL patients (including 33 children younger than 1 year of age) and 15 samples from AML patients (including 10 from children younger than 1 year of age). The main partner genes were
AFF1
(49%),
MLLT1
(27%),
MLLT3
(12%), and
MLLT10
(12%) in ALL and
MLLT3
(80%),
MLLT10
(10%), and
MLLT4
(10%) in AML. Fusion gene transcripts most commonly included
MLL
exon 11 (58% of ALL cases and 50% of AML cases), suggesting a breakpoint in
MLL
intron 11. |
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ISSN: | 0026-8933 1608-3245 |
DOI: | 10.1134/S0026893316060145 |