Investigation of the depletion of CARMEN lncRNA using a hiPSC model
Description
(1) We have obtained homozygous and heterozygous clones of the CARMEN-depleted human induced pluripotent stem cell (hiPSC) strain (Fig. 1). Depletion is achieved by insertion of polyadenylation poly(A) sites immediately behind the transcription start site of CARMEN gene. (2) Our preliminary data indicates that CARMEN depletion causes significant defect in cardiomyocyte differentiation (Fig. 2). The small percentage of successfully differentiated cells do have sarcomere structure (Fig.3), but the Z-disc patterning seems to be defective. (3) We have observed that there are two segments of the annotated CARMEN lncRNA (Fig. 4). And insertion of the poly(A) sequences perturbs the first halve (Fig. 5) but not the second halve (Fig. 6). (4) we investigated how CARMEN regulates target genomic regions, which were identified using computational approach by sequence homology. Depletion of the first halves of CARMEN didn’t significantly impact the expression of DNMT3A, TET3, TOP1 or WNT2B during iPSC-CM differentiation.