Generation of a retinoblastoma (Rb)1-inducible dominant-negative (DN) mouse model

Shikha Tarang, Songila M S R Doi, Channabasavaiah B Gurumurthy, Donald Harms, Rolen Quadros, Sonia M. Rocha-Sanchez

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Retinoblastoma 1 (Rb1) is an essential gene regulating cellular proliferation, differentiation, and homeostasis. To exert these functions, Rb1 is recruited and physically interacts with a growing variety of signaling pathways. While Rb1 does not appear to be ubiquitously expressed, its expression has been confirmed in a variety of hematopoietic and neuronalderived cells, including the inner ear hair cells (HCs). Studies in transgenic mice demonstrate that complete germline or conditional Rb1 deletion leads to abnormal cell proliferation, followed by massive apoptosis; making it difficult to fully address Rb1’s biochemical activities.To overcome these limitations, we developed a tetracycline-inducible TetO-CB-myc6-Rb1 (CBRb) mouse model to achieve transient and inducible dominantnegative (DN) inhibition of the endogenous RB1 protein. Our strategy involved fusing the Rb1 gene to the lysosomal protease pre-procathepsin B (CB), thus allowing for further routing of the DN-CBRb fusion protein and its interacting complexes for proteolytic degradation. Moreover, reversibility of the system is achieved upon suppression of doxycycline (Dox) administration. Preliminary characterization of DN-CBRb mice bred to a ubiquitous rtTA mouse line demonstrated a significant inhibition of the endogenous RB1 protein in the inner ear and in a number of other organs where RB1 is expressed. Examination of the postnatal (P) DN-CBRb mice inner ear at P10 and P28 showed the presence of supernumerary inner HCs (IHCs) in the lower turns of the cochleae, which corresponds to the described expression domain of the endogenous Rb1 gene. Selective and reversible suppression of gene expression is both an experimental tool for defining function and a potential means to medical therapy. Given the limitations associated with Rb1-null mice lethality, this model provides a valuable resource for understanding RB1 activity, relative contribution to HC regeneration and its potential therapeutic application.

Original languageEnglish (US)
JournalFrontiers in Cellular Neuroscience
Volume9
Issue numberFEB
DOIs
StatePublished - Feb 23 2015

Fingerprint

Retinoblastoma
Inner Auditory Hair Cells
Inner Ear
Retinoblastoma Genes
Cell Proliferation
Proteins
Doxycycline
Cochlea
Essential Genes
Tetracycline
Hair
Transgenic Mice
Regeneration
Homeostasis
Peptide Hydrolases
Apoptosis
Gene Expression
Therapeutics

Keywords

  • Dominant-negative
  • Doxycycline
  • Hair cell
  • Inner ear
  • Pre-procathepsin B
  • Regeneration
  • Retinoblastoma

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

Cite this

Generation of a retinoblastoma (Rb)1-inducible dominant-negative (DN) mouse model. / Tarang, Shikha; Doi, Songila M S R; Gurumurthy, Channabasavaiah B; Harms, Donald; Quadros, Rolen; Rocha-Sanchez, Sonia M.

In: Frontiers in Cellular Neuroscience, Vol. 9, No. FEB, 23.02.2015.

Research output: Contribution to journalArticle

Tarang, Shikha ; Doi, Songila M S R ; Gurumurthy, Channabasavaiah B ; Harms, Donald ; Quadros, Rolen ; Rocha-Sanchez, Sonia M. / Generation of a retinoblastoma (Rb)1-inducible dominant-negative (DN) mouse model. In: Frontiers in Cellular Neuroscience. 2015 ; Vol. 9, No. FEB.
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