Abstract
Organoid culture is a popular model to study gene function as the easy manipulating and time saving compared with in vivo experiments. This is widely used in auditory system for studying supporting cells (SCs) or hair cells (HCs) as only very few SCs or HCs can be harvested in both human and murine cochlea. However, the use of organoids is still a challenge due to the low efficiency in genetic modification. Here we took Lin28b as an example and compared Lin28b gain-of-function (GOF) and loss-of-function (LOF) with different genetic engineering methods and found that TetOn induced GOF or LOF was more efficient compared with lipofection or lentiviral transduction in the experimental conditions we used. Cell apoptosis in TetOn induction system was lowest compared with the other methods in this study. Our study is the first to compare the efficiency of different genetic engineering techniques in cochlear organoid culture, which may also apply to organoids established with other tissues.