Poster Presentation 47th Lorne Genome Conference 2026

Dynamic transcriptome-scale RNA-targeting with Cas13 (133384)

Dáire Gannon 1 2 3 , Maede Salehi 1 3 4 5 , Oliver Ozaydin 1 2 3 , Honglin Chen 6 7 , Mohamed Fareh 6 7 , Conor Kearney 4 5 , Stephin J Vervoort 1 2
  1. Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia
  2. Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia
  3. *, These authors contributed equally
  4. Olivia Newton-John Cancer Research Institute, Heidelberg, Victoria, Australia
  5. School of Cancer Medicine, La Trobe University, Melbourne, Victoria, Australia
  6. Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia
  7. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia

Cas13 enables rapid and reversible knockdown of RNA targets. However, to date the application of Cas13 to genome/transcriptome-scale high-throughput screening has been limited. Here, we have generated a transcriptome-scale gRNA library with >160,000 gRNAs targeting 19,000 coding and >8,000 non-coding RNAs, making it to our knowledge, the largest Cas13 gRNA library available. We performed Cas13 screens to identify essential coding transcripts in a range of cancer cell types and found a high concordance when benchmarking against publicly available RNAi and Cas9 screening data. We also found known essential non-coding RNAs as being essential in our Cas13 screens. Furthermore, we have performed a range of positive selection screens using clinically relevant immunological cell surface markers and drug resistance as readouts and have identified known and novel regulators of these pathways. Together, this data highlights the feasibility and utility of performing transcriptome-scale screens with Cas13.