Novel SF3B1 in-frame deletions result in aberrant RNA splicing in CLL patients
Blood Advances, 2017•Elsevier
Methods RNA-seq from 215 CLL patients was analyzed to discover if any novel genomic
abnormality was able to induce SF3B1-mutant-like aberrant splicing. Genomic abnormalities
in 3 aberrant splicing cases were identified as SF3B1 in-frame deletions and confirmed in
DNA by targeted resequencing. Analysis of RNA splicing was carried out as described
previously. 9 Expression of hemagglutinin (HA)-tagged mxSF3B1 deletion complementary
DNAs transfected into HEK293FT cells was confirmed by quantitative polymerase chain …
abnormality was able to induce SF3B1-mutant-like aberrant splicing. Genomic abnormalities
in 3 aberrant splicing cases were identified as SF3B1 in-frame deletions and confirmed in
DNA by targeted resequencing. Analysis of RNA splicing was carried out as described
previously. 9 Expression of hemagglutinin (HA)-tagged mxSF3B1 deletion complementary
DNAs transfected into HEK293FT cells was confirmed by quantitative polymerase chain …
Methods
RNA-seq from 215 CLL patients was analyzed to discover if any novel genomic abnormality was able to induce SF3B1-mutant-like aberrant splicing. Genomic abnormalities in 3 aberrant splicing cases were identified as SF3B1 in-frame deletions and confirmed in DNA by targeted resequencing. Analysis of RNA splicing was carried out as described previously. 9 Expression of hemagglutinin (HA)-tagged mxSF3B1 deletion complementary DNAs transfected into HEK293FT cells was confirmed by quantitative polymerase chain reaction (qPCR) and immunoblotting. Aberrant splicing was validated using ZDHHC16 minigenes and viability effect of E7107 in CLL primary cells was assessed by 3-(4, 5 dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. Methods in detail can be found in the supplemental Data.
Elsevier