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Inducible and Reversible Dominant-negative (DN) Protein Inhibition
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Genetics
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JoVE Journal Genetics
Inducible and Reversible Dominant-negative (DN) Protein Inhibition

Inducible and Reversible Dominant-negative (DN) Protein Inhibition

DOI:
10.3791/58419-v

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08:35 min

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January 07, 2019

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, , , ,
1Department of Oral Biology,Creighton University School of Dentistry

Chapters

  • 00:04Title
  • 00:58In Vitro Testing of the TetO-DN-CB-myc6-Rb1 Transgene in NIH3T3 Cells
  • 02:19In Vitro Testing of the TetO-DN-CB-myc6-Rb1 Transgene in HEI-OC1 Cells
  • 05:37Generation of CBRb+/ROSA-CAG-rtTA+ (CBRb) Transgenic Mice and In Vivo Testing of the DN-CBRb Approach
  • 06:09Results: Confirmation of Efficiency of TetO-DN-CB-myc6-Rb1 Construct
  • 07:50Conclusion

Summary

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Here we present a protocol to develop a dominant-negative inducible system, in which any protein can be conditionally inactivated by reversibly overexpressing a dominant-negative mutant version of it.

Tags

Dominant-negative ProteinConditional InactivationTransient InactivationPartial AblationEndogenous ExpressionMonomeric ProteinsNIH3T3 CellsPTet-SplicePCMV-Tet3GLipid-based TransfectionHEI-OC1 CellsCell ProliferationDoxycycline Induction

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