BRaf signaling principles unveiled by large-scale human mutation analysis with a rapid lentivirus-based gene replacement method

Chae Seok Lim, Xi Kang, Vincent Mirabella, Huaye Zhang, Qian Bu, Yoichi Araki, Elizabeth T. Hoang, Shiqiang Wang, Ying Shen, Sukwoo Choi, Bong Kiun Kaang, Qiang Chang, Zhiping P. Pang, Richard L. Huganir, J. Julius Zhu

Research output: Contribution to journalArticle

Abstract

Rapid advances in genetics are linking mutations on genes to diseases at an exponential rate, yet characterizing the gene mutation-cell behavior relationships essential for precision medicine remains a daunting task. More than 350 mutations on small GTPase BRaf are associated with various tumors, and ∼40 mutations are associated with the neurodevelopmental disorder cardio-facio-cutaneous syndrome (CFC). We developed a fast cost-effective lentivirus-based rapid gene replacement method to interrogate the physiopathology of BRaf and ∼50 disease-linked BRaf mutants, including all CFC-linked mutants. Analysis of simultaneous multiple patch-clamp recordings from 6068 pairs of rat neurons with validation in additional mouse and human neurons and multiple learning tests from 1486 rats identified BRaf as the key missing signaling effector in the common synaptic NMDA-R-CaMKII-SynGap-Ras-BRaf-MEK-ERK transduction cascade. Moreover, the analysis creates the original big data unveiling three general features of BRaf signaling. This study establishes the first efficient procedure that permits large-scale functional analysis of human disease-linked mutations essential for precision medicine.

Original languageEnglish (US)
Pages (from-to)537-552
Number of pages16
JournalGenes & development
Volume31
Issue number6
DOIs
StatePublished - Mar 15 2017

Keywords

  • BRaf
  • CFC syndrome
  • cancer
  • neurodevelopmental disorder
  • synaptic transmission

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology

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  • Cite this

    Lim, C. S., Kang, X., Mirabella, V., Zhang, H., Bu, Q., Araki, Y., Hoang, E. T., Wang, S., Shen, Y., Choi, S., Kaang, B. K., Chang, Q., Pang, Z. P., Huganir, R. L., & Zhu, J. J. (2017). BRaf signaling principles unveiled by large-scale human mutation analysis with a rapid lentivirus-based gene replacement method. Genes & development, 31(6), 537-552. https://doi.org/10.1101/gad.294413.116