Extended Data Fig. 7: Overview of sequencing analyses of in vivo screens using BSJ-gRNA libraries targeting 2,908 circRNAs.

a, Construction of the gRNA library targeting 2,908 circRNAs. One paired control gRNA (n = 2,908) and three BSJ-gRNAs (circRNA gRNAs, n = 8,724) were designed for each candidate circRNA. b, Cumulative distribution of the number of reads per gRNA of constructed libraries. The red line indicates that less than 1% of gRNAs are covered by less than 100 reads. c, Representation of 2,908 candidate circRNAs in different human tissues⁴¹ constructed in the library. On average, over 90% of top 100 abundant circRNAs in each tissue were included in the list of 2,908 candidate circRNAs. d, In vivo screen of circRNAs important for cell growth and proliferation. The gRNA lentiviral library was individually delivered into HT29 cells stably expressing RfxCas13d. Infected cells were enriched after 7 days and injected subcutaneously to nude mouse for 22 days. Genomic DNAs from infected cells were extracted at day 1 (D1) and 30 (D30) for gRNA amplification and deep sequencing. e, The Pearson correlation coefficient (PCC) between replicates (rep) of D1 and D30 in vivo samples in HT29. Two biologically independent experiments were performed at D1 and D30. f, Scatter plot of fold change of paired controls and circRNA BSJ-gRNAs between D1 and D30 in vivo samples in HT29. The grey dashed lines indicate 2 or 0.5 fold change, respectively. g, Analysis of gRNAs in FC ≤ 0.5 that identifies the same circRNA as a target gene. The number of circRNA candidates (n = 79), the average total gRNAs and altered gRNAs in FC ≤ 0.5 (purple) in in vivo screen are shown. Data are presented as means values ± s.d.