In vivo gain-of-function cDNA library screening for colonization genes in a mouse model of pulmonary metastasis

Zhenbo Tu; Antoine E Karnoub

doi:10.1016/j.xpro.2022.101413

In vivo gain-of-function cDNA library screening for colonization genes in a mouse model of pulmonary metastasis

STAR Protoc. 2022 May 25;3(2):101413. doi: 10.1016/j.xpro.2022.101413. eCollection 2022 Jun 17.

Authors

Zhenbo Tu¹, Antoine E Karnoub^{1

2

3}

Affiliations

¹ Department of Pathology and Cancer Center, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USA.
² Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
³ Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.

Abstract

We provide a protocol for gain-of-function (GOF) cDNA screen of genes that foster cancer cell colonization of secondary tissues, the last and most lethal step of the metastasis cascade. We present techniques for cDNA viral library preparation and delivery leading up to the recovery of colonization-promoting sequences in a proof-of-concept DU145-based mouse model of pulmonary metastasis. Adapted to other cDNA libraries and cancer models, this approach would prove widely useful in enumerating intrinsic genetic determinants underlying metastatic colonization. For complete details on the use and execution of this protocol, please refer to Tu et al. (2021).

Keywords: Cancer; Cell Biology; Gene Expression; Metabolism; Microscopy; Model Organisms; Molecular Biology; Molecular/Chemical Probes.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
DNA, Complementary / genetics
Disease Models, Animal
Gain of Function Mutation*
Gene Library
Lung Neoplasms* / genetics
Mice

Substances

DNA, Complementary

Grants and funding

R01 CA207322/CA/NCI NIH HHS/United States