Hargadon Kristian M, Bushhouse David Z, Johnson Coleman E, Williams Corey J
Hargadon Laboratory, Department of Biology, Hampden-Sydney College, Hampden-Sydney, VA, USA.
Methods Mol Biol. 2021;2265:25-46. doi: 10.1007/978-1-0716-1205-7_2.
Recent advances in the treatment of metastatic melanoma have emerged only from advances in our understanding of melanoma development and progression at the cellular and molecular levels. Despite the impact that such advances have made on the clinical management of this cancer over the last decade, additional insights into factors that promote melanoma progression and therapeutic resistance are needed to combat this disease. CRISPR-Cas9 gene editing technology is a powerful tool for studying gene function in a timely and cost-effective manner, enabling the manipulation of specific DNA sequences via a targeted approach. Herein, we describe a protocol for generating functional gene knockouts in melanoma cell lines by CRISPR-Cas9 gene editing, and we present an example application of this protocol for the successful knockout of the Foxc2 transcription factor-encoding gene in the B16-F1 murine melanoma cell line.
转移性黑色素瘤治疗的最新进展仅源于我们在细胞和分子水平上对黑色素瘤发生发展的认识进展。尽管这些进展在过去十年对这种癌症的临床管理产生了影响，但仍需要对促进黑色素瘤进展和治疗耐药性的因素有更多深入了解，以对抗这种疾病。CRISPR-Cas9基因编辑技术是一种以及时且经济高效的方式研究基因功能的强大工具，能够通过靶向方法操纵特定DNA序列。在此，我们描述了一种通过CRISPR-Cas9基因编辑在黑色素瘤细胞系中产生功能性基因敲除的方案，并展示了该方案在成功敲除B16-F1小鼠黑色素瘤细胞系中编码Foxc2转录因子的基因方面的示例应用。
