ena-STUDY-ETH ZURICH D-BSSE-03-11-2020-09:12:08:876-140
HEK293 cells transfected with polychromatic reporter constructs (pJD49 or pKB01) and respective inducer (PIT, ET or tTA) to identify/confirm the splicing patterns obtained.
The study was aimed at identifying the splice products obtained from the alternative promoter and alternative splicing based polychromatic reporter constructs upon induction of respective promoters. The use of fluorescent proteins as a read-out already gave an idea on the kind of splicing pattern to expect. However, the mRNA level data additionally confirms the outcome and sheds light on the mRNAs that are otherwise not visible. The initial construct, pKB01, did not express two of the three fluorescent proteins. Few sequence transitions led to improvements, making pJD49 that expressed all three fluorescent proteins. The study analyses cytoplasmic RNA obtained from transfected HEK293 cells for identifying the splicing pattern. We observe that splicing pattern changes from pKB01 to pJD49. Correctly spliced mRNA increases in pJD49, however, the mis-spliced mRNA levels still remain high in both pKB01 and pJD49.
The study was aimed at identifying the splice products obtained from the alternative promoter and alternative splicing based polychromatic reporter constructs upon induction of respective promoters. The use of fluorescent proteins as a read-out already gave an idea on the kind of splicing pattern to expect. However, the mRNA level data additionally confirms the outcome and sheds light on the mRNAs that are otherwise not visible. The initial construct, pKB01, did not express two of the three fluorescent proteins. Few sequence transitions led to improvements, making pJD49 that expressed all three fluorescent proteins. The study analyses cytoplasmic RNA obtained from transfected HEK293 cells for identifying the splicing pattern. We observe that splicing pattern changes from pKB01 to pJD49. Correctly spliced mRNA increases in pJD49, however, the mis-spliced mRNA levels still remain high in both pKB01 and pJD49.