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

Genes Dev. 2017 Mar 15;31(6):537-552. doi: 10.1101/gad.294413.116.


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.

Keywords: BRaf; CFC syndrome; cancer; neurodevelopmental disorder; synaptic transmission.

Publication types

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

MeSH terms

  • Animals
  • Cells, Cultured
  • Disease / genetics
  • Female
  • Gene Transfer Techniques
  • Humans
  • Lentivirus / genetics
  • MAP Kinase Signaling System / genetics*
  • Male
  • Mice, Inbred C57BL
  • Mutation*
  • Neurons / physiology
  • Proto-Oncogene Proteins B-raf / genetics*
  • Rats, Sprague-Dawley
  • Synaptic Transmission / genetics*
  • Tissue Culture Techniques


  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf