Doublecortin-like kinase 1 (DCLK1) is critical for neurogenesis, but overexpression is also observed in multiple cancers and is associated with poor prognosis. Nevertheless, the function of DCLK1 in cancer, especially the context-dependent functions, are poorly understood. We present a “toolkit” that includes the DCLK1 inhibitor DCLK1-IN-1, a complementary DCLK1-IN-1-resistant mutation G532A, and kinase dead mutants D511N and D533N, which can be used to investigate signaling pathways regulated by DCLK1. Using a cancer cell line engineered to be DCLK1 dependent for growth and cell migration, we show that this toolkit can be used to discover associations between DCLK1 kinase activity and biological processes. In particular, we show an association between DCLK1 and RNA processing, including the identification of CDK11 as a potential substrate of DCLK1 using phosphoproteomics.

Doublecortin样激酶1（DCLK1）对于神经发生至关重要，但在多种癌症中也观察到过表达，并且与不良预后相关。然而，人们对DCLK1在癌症中的功能，特别是与上下文相关的功能知之甚少。我们提出了一个“工具包”，其中包括DCLK1抑制剂DCLK1-IN-1，互补的DCLK1-IN-1抗性突变G532A，以及激酶死亡突变体D511N和D533N，可用于研究DCLK1调控的信号通路。使用被设计成依赖于DCLK1的癌细胞系进行生长和细胞迁移，我们表明该工具包可用于发现DCLK1激酶活性与生物学过程之间的关联。特别是，我们展示了DCLK1与RNA处理之间的关联，