Conditional gene silencing via a CRISPR system in cerebellar Purkinje cells

• Published in: Biochimica et biophysica acta. General subjects Vol. 1865; no. 6; p. 129869
• Main Authors: Wu, Qin-Wei, Kapfhammer, Josef P.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.06.2021
• Subjects: Animals; cerebellum; Cerebellum - metabolism; Cerebellum - pathology; CRISPR-Cas Systems; CRISPR-Cas13; Gene Silencing; gene targeting; genes; Mice; Mice, Knockout; neurons; Protein Kinase C - antagonists & inhibitors; Protein Kinase C - genetics; protein synthesis; Purkinje cells; Purkinje Cells - metabolism; Purkinje Cells - pathology; CRISPR-Cas13; gene silencing; Purkinje cells
• ISSN: 0304-4165; 1872-8006; 1872-8006
• DOI: 10.1016/j.bbagen.2021.129869

•Specific gene knockdown in cerebellar Purkinje cells is a major challenge because Purkinje cells only make up a small fraction of the cerebellum and off-target effects of shRNA occur.•Little is known about the application of CRISPR-Cas13 and the resulting protein expression levels in Purkinje cells, a type of developing postmitotic neuron.•We explored the possibility of a Cas13-mediated conditional knockdown system for cerebellar Purkinje cells. This system can achieve a suppression of the target proteins restricted to Purkinje cells in mixed cerebellar cultures.