Forward and Reverse Genetics of B Cell Malignancies: From Insertional Mutagenesis to CRISPR-Cas
Cancer genome sequencing has identified dozens of mutations with a putative role in lymphomagenesis and leukemogenesis. Validation of driver mutations responsible for B cell neoplasms is complicated by the volume of mutations worthy of investigation and by the complex ways that multiple mutations ar...
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doaj-6e78295678eb4b21a0edfe05fac25f662021-08-13T08:46:55ZengFrontiers Media S.A.Frontiers in Immunology1664-32242021-08-011210.3389/fimmu.2021.670280670280Forward and Reverse Genetics of B Cell Malignancies: From Insertional Mutagenesis to CRISPR-CasJoanna C. Dawes0Joanna C. Dawes1Anthony G. Uren2Anthony G. Uren3Medical Research Council, London Institute of Medical Sciences, London, United KingdomInstitute of Clinical Sciences (ICS), Faculty of Medicine, Imperial College London, London, United KingdomMedical Research Council, London Institute of Medical Sciences, London, United KingdomInstitute of Clinical Sciences (ICS), Faculty of Medicine, Imperial College London, London, United KingdomCancer genome sequencing has identified dozens of mutations with a putative role in lymphomagenesis and leukemogenesis. Validation of driver mutations responsible for B cell neoplasms is complicated by the volume of mutations worthy of investigation and by the complex ways that multiple mutations arising from different stages of B cell development can cooperate. Forward and reverse genetic strategies in mice can provide complementary validation of human driver genes and in some cases comparative genomics of these models with human tumors has directed the identification of new drivers in human malignancies. We review a collection of forward genetic screens performed using insertional mutagenesis, chemical mutagenesis and exome sequencing and discuss how the high coverage of subclonal mutations in insertional mutagenesis screens can identify cooperating mutations at rates not possible using human tumor genomes. We also compare a set of independently conducted screens from Pax5 mutant mice that converge upon a common set of mutations observed in human acute lymphoblastic leukemia (ALL). We also discuss reverse genetic models and screens that use CRISPR-Cas, ORFs and shRNAs to provide high throughput in vivo proof of oncogenic function, with an emphasis on models using adoptive transfer of ex vivo cultured cells. Finally, we summarize mouse models that offer temporal regulation of candidate genes in an in vivo setting to demonstrate the potential of their encoded proteins as therapeutic targets.https://www.frontiersin.org/articles/10.3389/fimmu.2021.670280/fullB cell leukemiaB cell lymphomamouse modelsinsertional mutagenesisexome sequencingreverse genetics |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Joanna C. Dawes Joanna C. Dawes Anthony G. Uren Anthony G. Uren |
spellingShingle |
Joanna C. Dawes Joanna C. Dawes Anthony G. Uren Anthony G. Uren Forward and Reverse Genetics of B Cell Malignancies: From Insertional Mutagenesis to CRISPR-Cas Frontiers in Immunology B cell leukemia B cell lymphoma mouse models insertional mutagenesis exome sequencing reverse genetics |
author_facet |
Joanna C. Dawes Joanna C. Dawes Anthony G. Uren Anthony G. Uren |
author_sort |
Joanna C. Dawes |
title |
Forward and Reverse Genetics of B Cell Malignancies: From Insertional Mutagenesis to CRISPR-Cas |
title_short |
Forward and Reverse Genetics of B Cell Malignancies: From Insertional Mutagenesis to CRISPR-Cas |
title_full |
Forward and Reverse Genetics of B Cell Malignancies: From Insertional Mutagenesis to CRISPR-Cas |
title_fullStr |
Forward and Reverse Genetics of B Cell Malignancies: From Insertional Mutagenesis to CRISPR-Cas |
title_full_unstemmed |
Forward and Reverse Genetics of B Cell Malignancies: From Insertional Mutagenesis to CRISPR-Cas |
title_sort |
forward and reverse genetics of b cell malignancies: from insertional mutagenesis to crispr-cas |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Immunology |
issn |
1664-3224 |
publishDate |
2021-08-01 |
description |
Cancer genome sequencing has identified dozens of mutations with a putative role in lymphomagenesis and leukemogenesis. Validation of driver mutations responsible for B cell neoplasms is complicated by the volume of mutations worthy of investigation and by the complex ways that multiple mutations arising from different stages of B cell development can cooperate. Forward and reverse genetic strategies in mice can provide complementary validation of human driver genes and in some cases comparative genomics of these models with human tumors has directed the identification of new drivers in human malignancies. We review a collection of forward genetic screens performed using insertional mutagenesis, chemical mutagenesis and exome sequencing and discuss how the high coverage of subclonal mutations in insertional mutagenesis screens can identify cooperating mutations at rates not possible using human tumor genomes. We also compare a set of independently conducted screens from Pax5 mutant mice that converge upon a common set of mutations observed in human acute lymphoblastic leukemia (ALL). We also discuss reverse genetic models and screens that use CRISPR-Cas, ORFs and shRNAs to provide high throughput in vivo proof of oncogenic function, with an emphasis on models using adoptive transfer of ex vivo cultured cells. Finally, we summarize mouse models that offer temporal regulation of candidate genes in an in vivo setting to demonstrate the potential of their encoded proteins as therapeutic targets. |
topic |
B cell leukemia B cell lymphoma mouse models insertional mutagenesis exome sequencing reverse genetics |
url |
https://www.frontiersin.org/articles/10.3389/fimmu.2021.670280/full |
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