Cargando…

piggyBac Transposon Somatic Mutagenesis with an Activated Reporter and Tracker (PB-SMART) for Genetic Screens in Mice

Somatic forward genetic screens have the power to interrogate thousands of genes in a single animal. Retroviral and transposon mutagenesis systems in mice have been designed and deployed in somatic tissues for surveying hematopoietic and solid tumor formation. In the context of cancer, the ability t...

Descripción completa

Detalles Bibliográficos
Autores principales: Landrette, Sean F., Cornett, Jonathan C., Ni, Thomas K., Bosenberg, Marcus W., Xu, Tian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3198810/
https://www.ncbi.nlm.nih.gov/pubmed/22039523
http://dx.doi.org/10.1371/journal.pone.0026650
_version_ 1782214495781781504
author Landrette, Sean F.
Cornett, Jonathan C.
Ni, Thomas K.
Bosenberg, Marcus W.
Xu, Tian
author_facet Landrette, Sean F.
Cornett, Jonathan C.
Ni, Thomas K.
Bosenberg, Marcus W.
Xu, Tian
author_sort Landrette, Sean F.
collection PubMed
description Somatic forward genetic screens have the power to interrogate thousands of genes in a single animal. Retroviral and transposon mutagenesis systems in mice have been designed and deployed in somatic tissues for surveying hematopoietic and solid tumor formation. In the context of cancer, the ability to visually mark mutant cells would present tremendous advantages for identifying tumor formation, monitoring tumor growth over time, and tracking tumor infiltrations and metastases into wild-type tissues. Furthermore, locating mutant clones is a prerequisite for screening and analyzing most other somatic phenotypes. For this purpose, we developed a system using the piggyBac (PB) transposon for somatic mutagenesis with an activated reporter and tracker, called PB-SMART. The PB-SMART mouse genetic screening system can simultaneously induce somatic mutations and mark mutated cells using bioluminescence or fluorescence. The marking of mutant cells enable analyses that are not possible with current somatic mutagenesis systems, such as tracking cell proliferation and tumor growth, detecting tumor cell infiltrations, and reporting tissue mutagenesis levels by a simple ex vivo visual readout. We demonstrate that PB-SMART is highly mutagenic, capable of tumor induction with low copy transposons, which facilitates the mapping and identification of causative insertions. We further integrated a conditional transposase with the PB-SMART system, permitting tissue-specific mutagenesis with a single cross to any available Cre line. Targeting the germline, the system could also be used to conduct F1 screens. With these features, PB-SMART provides an integrated platform for individual investigators to harness the power of somatic mutagenesis and phenotypic screens to decipher the genetic basis of mammalian biology and disease.
format Online
Article
Text
id pubmed-3198810
institution National Center for Biotechnology Information
language English
publishDate 2011
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-31988102011-10-28 piggyBac Transposon Somatic Mutagenesis with an Activated Reporter and Tracker (PB-SMART) for Genetic Screens in Mice Landrette, Sean F. Cornett, Jonathan C. Ni, Thomas K. Bosenberg, Marcus W. Xu, Tian PLoS One Research Article Somatic forward genetic screens have the power to interrogate thousands of genes in a single animal. Retroviral and transposon mutagenesis systems in mice have been designed and deployed in somatic tissues for surveying hematopoietic and solid tumor formation. In the context of cancer, the ability to visually mark mutant cells would present tremendous advantages for identifying tumor formation, monitoring tumor growth over time, and tracking tumor infiltrations and metastases into wild-type tissues. Furthermore, locating mutant clones is a prerequisite for screening and analyzing most other somatic phenotypes. For this purpose, we developed a system using the piggyBac (PB) transposon for somatic mutagenesis with an activated reporter and tracker, called PB-SMART. The PB-SMART mouse genetic screening system can simultaneously induce somatic mutations and mark mutated cells using bioluminescence or fluorescence. The marking of mutant cells enable analyses that are not possible with current somatic mutagenesis systems, such as tracking cell proliferation and tumor growth, detecting tumor cell infiltrations, and reporting tissue mutagenesis levels by a simple ex vivo visual readout. We demonstrate that PB-SMART is highly mutagenic, capable of tumor induction with low copy transposons, which facilitates the mapping and identification of causative insertions. We further integrated a conditional transposase with the PB-SMART system, permitting tissue-specific mutagenesis with a single cross to any available Cre line. Targeting the germline, the system could also be used to conduct F1 screens. With these features, PB-SMART provides an integrated platform for individual investigators to harness the power of somatic mutagenesis and phenotypic screens to decipher the genetic basis of mammalian biology and disease. Public Library of Science 2011-10-21 /pmc/articles/PMC3198810/ /pubmed/22039523 http://dx.doi.org/10.1371/journal.pone.0026650 Text en Landrette et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Landrette, Sean F.
Cornett, Jonathan C.
Ni, Thomas K.
Bosenberg, Marcus W.
Xu, Tian
piggyBac Transposon Somatic Mutagenesis with an Activated Reporter and Tracker (PB-SMART) for Genetic Screens in Mice
title piggyBac Transposon Somatic Mutagenesis with an Activated Reporter and Tracker (PB-SMART) for Genetic Screens in Mice
title_full piggyBac Transposon Somatic Mutagenesis with an Activated Reporter and Tracker (PB-SMART) for Genetic Screens in Mice
title_fullStr piggyBac Transposon Somatic Mutagenesis with an Activated Reporter and Tracker (PB-SMART) for Genetic Screens in Mice
title_full_unstemmed piggyBac Transposon Somatic Mutagenesis with an Activated Reporter and Tracker (PB-SMART) for Genetic Screens in Mice
title_short piggyBac Transposon Somatic Mutagenesis with an Activated Reporter and Tracker (PB-SMART) for Genetic Screens in Mice
title_sort piggybac transposon somatic mutagenesis with an activated reporter and tracker (pb-smart) for genetic screens in mice
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3198810/
https://www.ncbi.nlm.nih.gov/pubmed/22039523
http://dx.doi.org/10.1371/journal.pone.0026650
work_keys_str_mv AT landretteseanf piggybactransposonsomaticmutagenesiswithanactivatedreporterandtrackerpbsmartforgeneticscreensinmice
AT cornettjonathanc piggybactransposonsomaticmutagenesiswithanactivatedreporterandtrackerpbsmartforgeneticscreensinmice
AT nithomask piggybactransposonsomaticmutagenesiswithanactivatedreporterandtrackerpbsmartforgeneticscreensinmice
AT bosenbergmarcusw piggybactransposonsomaticmutagenesiswithanactivatedreporterandtrackerpbsmartforgeneticscreensinmice
AT xutian piggybactransposonsomaticmutagenesiswithanactivatedreporterandtrackerpbsmartforgeneticscreensinmice