Newly, Dicer-substrate small interfering RNA (DsiRNA) has gathered interest due to its better efficacy in comparison to small interfering RNA (siRNA). Yet, DsiRNA’s use has been constrained due to its quick degradation. The micelles of cholic acid-polyethylenimine (CA-PEI) copolymer, as carriers of DsiRNA were developed in this study. CA is an amphiphilic steroid molecule made naturally from cholesterol, and has the potential to arrange itself into micelles. PEI is a cationic molecule mostly used to deliver genes because of its potential to condense nucleic acid, nuclear localization capability, endosome escaping ability and effective transfection ability. The mean particle size and zeta potential of DsiRNA-loaded CA-PEI micelles were studied and were discovered to be ranging from 125-150 nm, with zeta potential in the range of +4-+12 mV. The morphology of DsiRNA-loaded CA-PEI micelles was investigated under the transmission electron microscopy, and the micelles were found to be spherical but aggregated. The DsiRNA release from CA-PEI micelles was also studied using the dialysis membrane bags, and the results showed an initial burst release followed by a sustained release. The DsiRNA-loaded CA-PEI micelles had a high loading efficiency, and DsiRNA was strongly bound. The DsiRNA-loaded CA-PEI micelles were found to be nontoxic to the normal (V79) and cancer (DLD-1) cells. The study showed the potential of CA-PEI micelles as the promising carriers of DsiRNA.