Journal of the Association for Research in Otolaryngology

, Volume 13, Issue 3, pp 295–322

Conditional Gene Expression in the Mouse Inner Ear Using Cre-loxP

  • Brandon C. Cox
  • Zhiyong Liu
  • Marcia M. Mellado Lagarde
  • Jian Zuo
Invited Review

DOI: 10.1007/s10162-012-0324-5

Cite this article as:
Cox, B.C., Liu, Z., Lagarde, M.M.M. et al. JARO (2012) 13: 295. doi:10.1007/s10162-012-0324-5

Abstract

In recent years, there has been significant progress in the use of Cre-loxP technology for conditional gene expression in the inner ear. Here, we introduce the basic concepts of this powerful technology, emphasizing the differences between Cre and CreER. We describe the creation and Cre expression pattern of each Cre and CreER mouse line that has been reported to have expression in auditory and vestibular organs. We compare the Cre expression patterns between Atoh1-CreERTM and Atoh1-CreERT2 and report a new line, Fgfr3-iCreERT2, which displays inducible Cre activity in cochlear supporting cells. We also explain how results can vary when transgenic vs. knock-in Cre/CreER alleles are used to alter gene expression. We discuss practical issues that arise when using the Cre-loxP system, such as the use of proper controls, Cre efficiency, reporter expression efficiency, and Cre leakiness. Finally, we introduce other methods for conditional gene expression, including Flp recombinase and the tetracycline-inducible system, which can be combined with Cre-loxP mouse models to investigate conditional expression of more than one gene.

Keywords

cochleaconditional gene deletion CreERCre efficiencyCre recombinaseectopic gene expressionFlp recombinaseknock-inLoxPreporter linesTet-onTet-offtransgenicutriclevestibular

Abbreviations

ABR

Auditory brainstem response

BAC

Bacterial artificial chromosome

βgal

Beta-galactosidase

BMP

Bone morphogenetic protein

BDNF

Brain-derived neurotrophic factor

CFP

Cyan fluorescent protein

Col1A1

Alpha (1) collagen promoter

Col2A1

Type II collagen promoter

DTA

Diphtheria toxin fragment A

E

Embryonic day

eGFP

Enhanced green fluorescent protein

ES

Embryonic stem cells

GER

Greater epithelial ridge

GFAP

Glial fibrillary acidic protein

GFP

Green fluorescent protein

GOI

Gene of interest

Hsp90

Heat shock protein 90

iCsp3

Drug-inducible dimerizable caspase 3

IRES

Internal ribosome entry site

KOMP

NIH knockout mouse project

LER

Lesser epithelial ridge

myo7a

Myosin VIIa

NICD

Notch intracellular domain

P

Postnatal day

PAC

Phage artificial chromosome

Pkd1

Polycystic kidney disease 1

Rb

Retinoblastoma protein

RFP

Red fluorescent protein

rtTA

Reverse tetracycline transactivator

SHH

Sonic hedgehog

TRE

Tetracycline responsive element

tTA

Tetracycline transactivator

Copyright information

© Association for Research in Otolaryngology 2012

Authors and Affiliations

  • Brandon C. Cox
    • 1
  • Zhiyong Liu
    • 1
  • Marcia M. Mellado Lagarde
    • 1
  • Jian Zuo
    • 1
  1. 1.Department of Developmental NeurobiologySt. Jude Children’s Research HospitalMemphisUSA