Fig. 3: Screening in vivo using the Menuetto library.
A Diagram of the design of the whole-genome in vivo screen using the Menuetto library and FLCs derived from Eμ-MycT/+;enAsCas12aKI/+ crosses. B Survival curve of mice transplanted with Eμ-MycT/+;enAsCas12aKI/+ FLCs for in vivo screening with the Menuetto pre-crRNA library. crNTC mice remain mostly alive after 95 days, with only one mouse succumbing to pre-B/B cell lymphoma (n = 1/6). Two crNTC control mice have been excluded from the curve (original cohort n = 8) as their death was determined to have a cause other than pre-B/B cell lymphoma (via immunophenotyping). crTrp53 control mice (n = 5/5) reached ethical endpoint with a median latency of 25 days. Menuetto library mice (n = 17/21) have reached ethical endpoint with a median latency of 48 days. Mantel-Cox log-rank test comparisons: crNTC vs Menuetto p = 0.0174, crNTC vs crTrp53 p = 0.0008, crTrp53 vs Menuetto p < 0.0001. C Immunophenotyping results from the tumours of the 17 Menuetto library mice that have so far succumbed to lymphoma. D Graph of highly enriched pre-crRNAs in the haematopoietic tissue tumours of the 17 mice obtained so far from the in vivo Menuetto library screen. Trp53-targeting pre-crRNAs were the most common, and likely causative even in the instances where multiple, roughly equally represented pre-crRNAs were detected (Trp53 + other; n = 6). Source data are provided as a Source Data file. Abbreviations: crRNA CRISPR RNA, FLCs foetal liver cells, WT wild-type, NTC non-targeting control, NGS next-generation sequencing.
