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Blood, Vol. 110, Issue 1, 388-392, July 1, 2007
Different chromosomal breakpoints impact the level of LMO2 expression in T-ALL
Blood Dik et al.
110: 388
Supplemental materials for: Dik et al, Volume 110, Issue 1, 388-392
Files in this Data Supplement:
- Table S1. Functionality of LMO2 12bp-cRSS based on ex vivo recombination substrate assay and RIC algorithm (PDF, 26.1 KB)
- Table S2. 12 bp- and 23 bp-cRSSs in LMO2 that pass the RIC score (PDF, 30.7 KB)
- Figure S1. Recombination vector (schematic) (JPG, 49.7 KB)
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The LMO2 sequence of interest is cloned in an upstream MluI-SalI cassette. “Ptac” represents promoter; “CAT,” chloramphenicol acetyltransferase gene; and “stop,” transcriptional terminator. The authentic D3 RSS is shown as a triangle (a). cRSSs are represented by incomplete triangles: b, LMO2 cRSS observed near the t(11;14)(p13;q11) translocation breakpoint: c, other LMO2 cRSSs potentially present within the cloned area: d, cRSS within the core vector sequence. Recombination pathways: pathway 1, V(D)J recombination between the authentic D3 RSS and the LMO2 cRSS to be tested; 2, V(D)J recombination between the authentic D3 RSS and another cRSS potentially present within the cloned LMO2 part; 3, V(D)J recombination between the authentic D3 RSS and cRSS within the plasmid core sequence; and 4, aspecific/break repair recombination. All 4 pathways remove the transcription termination sequence with subsequent activation of the CAT gene, which is used as selection marker for plasmids having undergone recombination events.
- Figure S2. Breakpoint sequences and flanking sequences of derivative chromosomes of 9 T-ALL cases with t(11;14)(p13;q11) (JPG, 257 KB)
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Normal TCRD sequences on chromosome 14q11 are indicated (14 TCRD) as well as germline LMO2 sequences (11 LMO2). der(14) and der(11) indicate the sequences of the derivative chromosomes 14 and 11, respectively. TCRD gene segments are indicated, and nucleotides belonging to the heptamer and nonamer sequences of TCRD recombination signal sequences (RSSs) are underlined. Untemplated nucleotides (N-regions) are in italics. Putative cRSSs in the vicinity of the LMO2 break are bold and marked with a dotted line. Cases 647, T121, and T024 have a der(14) with a D3/LMO2 fusion and a der(11) with a fusion between LMO2 and the 3′ D3 23bp-RSS. Case T068, der(14) with a D2/LMO2 fusion. Case 6206, der(11) with a LMO2/J1 fusion. Case T064, der(11) with a LMO2/D2 fusion. Case UPN4395, der(14) with an inverted D2/LMO2 and der(11) with a LMO2/V3 fusion. Case UPN1589, der(14) with a D2/LMO2 and der(11) with a LMO2/J1 fusion; the boxed area in this case indicates an LMO2 duplication of 493 bp that is present on both derivative chromosomes. TALL-104, der(14) with a D2/LMO2 and der(11) with a LMO2/D3 fusion.
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