Suddhasil Mookherjee

Mutations in CEP290 cause ciliogenesis defects, leading to diverse clinical phenotypes, including Leber congenital amaurosis (LCA). Gene therapy for CEP290-associated diseases is hindered by the 7.4 kb CEP290 coding sequence, which is difficult to deliver in vivo. The multi-domain structure of the CEP290 protein suggests that a specific CEP290 domain may complement disease phenotypes.

Expression cassettes of SpCas9 and sgRNA were delivered by two separate AAV vectors (Fig. 1a). The Streptococcus pyogenes Cas9 (SpCas9) coding sequence was placed in between a photoreceptor-specific human rhodopsin kinase (RK) promoter20 and a synthetic polyadenylation signal. A human RNA polymerase III promoter U6 was used to drive sgRNA expression. A tdTomato expression cassette was included in the sgRNA vector to track transduction (Fig. 1a).