Generation of a CRISPR activation mouse that enables modelling of aggressive lymphoma and interrogation of venetoclax resistance.
Yexuan DengSarah T DiepstratenMargaret A PottsGöknur GinerStephanie TreziseAshley P NgGerry HealeySerena R KaneAmali CoorayKira BehrensAmy HeidersbachAndrew J KuehMartin PalStephen WilcoxLin TaiWarren S AlexanderJane E VisvaderStephen L NuttAndreas StrasserBenjamin HaleyQuan ZhaoGemma L KellyMarco J HeroldPublished in: Nature communications (2022)
CRISPR technologies have advanced cancer modelling in mice, but CRISPR activation (CRISPRa) methods have not been exploited in this context. We establish a CRISPRa mouse (dCas9a-SAM KI ) for inducing gene expression in vivo and in vitro. Using dCas9a-SAM KI primary lymphocytes, we induce B cell restricted genes in T cells and vice versa, demonstrating the power of this system. There are limited models of aggressive double hit lymphoma. Therefore, we transactivate pro-survival BCL-2 in Eµ-Myc T/+ ;dCas9a-SAM KI/+ haematopoietic stem and progenitor cells. Mice transplanted with these cells rapidly develop lymphomas expressing high BCL-2 and MYC. Unlike standard Eµ-Myc lymphomas, BCL-2 expressing lymphomas are highly sensitive to the BCL-2 inhibitor venetoclax. We perform genome-wide activation screens in these lymphoma cells and find a dominant role for the BCL-2 protein A1 in venetoclax resistance. Here we show the potential of our CRISPRa model for mimicking disease and providing insights into resistance mechanisms towards targeted therapies.
Keyphrases
- genome wide
- dna methylation
- gene expression
- induced apoptosis
- advanced cancer
- diffuse large b cell lymphoma
- cell cycle arrest
- copy number
- palliative care
- genome editing
- crispr cas
- transcription factor
- neoadjuvant chemotherapy
- chronic lymphocytic leukemia
- signaling pathway
- high fat diet induced
- endoplasmic reticulum stress
- type diabetes
- cell death
- squamous cell carcinoma
- metabolic syndrome
- risk assessment
- cell proliferation
- mass spectrometry
- protein protein
- adipose tissue
- locally advanced
- anti inflammatory
- simultaneous determination
- free survival
- fluorescent probe
- tandem mass spectrometry