KCNJ11 knockout morula re-engineered by stem cell diploid aggregation

Timothy J. Nelson, Almudena Martinez-Fernandez, Andre Terzic

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

KCNJ11-encoded Kir6.2 assembles with ATP-binding cassette sulphonylurea receptors to generate ATP-sensitive K+ (KATP) channel complexes. Expressed in tissues with dynamic metabolic flux, these evolutionarily conserved yet structurally and functionally unique heteromultimers serve as high-fidelity rheostats that adjust membrane potential-dependent cell functions to match energetic demand. Genetic defects in channel subunits disrupt the cellular homeostatic response to environmental stress, compromising organ tolerance in the adult. As maladaptation characterizes malignant KATP channelopathies, establishment of platforms to examine progression of KATP channel-dependent adaptive behaviour is warranted. Chimeras provide a powerful tool to assay the contribution of genetic variance to stress intolerance during prenatal or post-natal development. Here, KCNJ11 KATP channel gene knockout ↔ wild-type chimeras were engineered through diploid aggregation. Integration of wild-type embryonic stem cells into zona pellucida-denuded morula derived from knockout embryos achieved varying degrees of incorporation of stress-tolerant tissue within the KATP channel-deficient background. Despite the stress-vulnerable phenotype of the knockout, ex vivo derived mosaic blastocysts tolerated intrauterine transfer and implantation, followed by full-term embryonic development in pseudopregnant surrogates to produce live chimeric offspring. The development of adult chimerism from the knockout ↔ wild-type mosaic embryo offers thereby a new paradigm to probe the ecogenetic control of the KATP channel-dependent stress response.

Original languageEnglish (US)
Pages (from-to)269-276
Number of pages8
JournalPhilosophical Transactions of the Royal Society B: Biological Sciences
Volume364
Issue number1514
DOIs
StatePublished - 2009

Keywords

  • ATP-sensitive K channel
  • Chimerism
  • Ecogenetics
  • Genetic variance
  • Kir6.2
  • stress

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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