The past and presence of gene targeting: from chemicals and DNA via proteins to RNA

T M Geel, M H J Ruiters, R H Cool, L Halby, D C Voshart, L Andrade Ruiz, K E Niezen-Koning, P B Arimondo, M G Rots

Research output: Contribution to journalReview articleAcademicpeer-review

9 Citations (Scopus)

Abstract

The ability to target DNA specifically at any given position within the genome allows many intriguing possibilities and has inspired scientists for decades. Early gene-targeting efforts exploited chemicals or DNA oligonucleotides to interfere with the DNA at a given location in order to inactivate a gene or to correct mutations. We here describe an example towards correcting a genetic mutation underlying Pompe's disease using a nucleotide-fused nuclease (TFO-MunI). In addition to the promise of gene correction, scientists soon realized that genes could be inactivated or even re-activated without inducing potentially harmful DNA damage by targeting transcriptional modulators to a particular gene. However, it proved difficult to fuse protein effector domains to the first generation of programmable DNA-binding agents. The engineering of gene-targeting proteins (zinc finger proteins (ZFPs), transcription activator-like effectors (TALEs)) circumvented this problem. The disadvantage of protein-based gene targeting is that a fusion protein needs to be engineered for every locus. The recent introduction of CRISPR/Cas offers a flexible approach to target a (fusion) protein to the locus of interest using cheap designer RNA molecules. Many research groups now exploit this platform and the first human clinical trials have been initiated: CRISPR/Cas has kicked off a new era of gene targeting and is revolutionizing biomedical sciences.This article is part of a discussion meeting issue 'Frontiers in epigenetic chemical biology'.

Original languageEnglish
Article number20170077
JournalPhilosophical Transactions of the Royal Society of London. Series B: Biological Sciences
Volume373
Issue number1748
Early online date23-Apr-2018
DOIs
Publication statusPublished - 5-Jun-2018

Keywords

  • METHYLATION
  • MODULATION
  • TRIPLEX-FORMING OLIGONUCLEOTIDES
  • PYRROLE-IMIDAZOLE POLYAMIDES
  • IN-VIVO
  • MINOR-GROOVE
  • HUMAN-CELLS
  • GENOME
  • TRANSCRIPTION
  • CANCER

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