Retinitis pigmentosa 11 (RP11) is an inherited retinal dystrophy caused by heterozygous mutations in pre-mRNA processing factor 31 (PRPF31). CCR4-NOT transcription complex subunit 3 (CNOT3), a transcriptional inhibitor of PRPF31, is found at higher levels in RP11 patients compared to asymptomatic family members carrying the same mutations. In this study, we aim to upregulate functional PRPF31 to enhance pre-mRNA processing and rescue disease phenotypes by lowering CNOT3 function. We found 10% higher expression of CNOT3 with 17% lower PRPF31 in a patient compared to an asymptomatic relative, both carrying heterozygous PRPF31 c.1205 C>A. Antisense oligomers were designed to target removal of CNOT3 exon(s) encoding essential functional domains, resulting in truncated CNOT3 isoform(s). Results exhibited 2-fold increase in PRPF31 expression in antisense oligomer treated RP11 iPSC-derived retinal pigment epithelium. Consequently, treated cells showed a significant increase in cilia incidence and length, factors crucial for retinal function. Poly(A) RT-PCR also revealed increase in PRPF31 containing poly(A)-tail in CNOT3 depleted cells. In conclusion, this study shows antisense oligomers can lower CNOT3 function and increase PRPF31 transcription and mRNA stability to levels expected to provide therapeutic benefit. Transcriptome analysis will be used to evaluate pre-mRNA splicing after increased PRPF31 expression.