Department of Pathology and Cancer Center, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USA.
Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
STAR Protoc. 2022 May 25;3(2):101413. doi: 10.1016/j.xpro.2022.101413. eCollection 2022 Jun 17.
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).
我们提供了一个功能获得（GOF）cDNA 筛选基因的方案，这些基因促进癌细胞在次级组织中的定植，这是转移级联的最后也是最致命的步骤。我们介绍了 cDNA 病毒文库制备和递送的技术，这些技术可在以 DU145 为基础的肺转移小鼠模型中回收促进定植的序列，从而证明该方案的可行性。该方法适用于其他 cDNA 文库和癌症模型，如果加以适当调整，将广泛用于确定转移定植的内在遗传决定因素。有关该方案使用和实施的完整详细信息，请参阅 Tu 等人（2021 年）。
