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Catalogue No.


Cell Line Name


Cell Line Description

This cell line can be used to determine the function of a protein by rapidly inactivating the protein of interest by rapamycin induced rerouting of the protein to the mitochondria. The HeLa-Mitotrap cell line is a stably transfected HeLa cell line expressing a mitochondrial trapping protein containing an FRB domain. The term FRB refers to a protein containing FKBP12 and Rapamycin Binding (FRB) domains within the mTOR (mammalian Target of Rapamycin) protein. The Hela-Mitotrap cell line can be transfected to express the target protein of interest genetically modified to have an FKBP domain. Upon treatment of the cells with rapamycin target proteins containing an FKBP domain will dimerize with FRB domains. The resulting protein complexes are sequestered to the mitochondria.Treatment of cell lines containing both FRB and FKBP rapamycin binding proteins can sequester target proteins to the mitochondria within minutes of treatment rendering the cells ready for immediate assay. This type of approach, referred to as ‘‘knocksideways,'' should be widely applicable as a means of inactivating proteins within a timescale of seconds or minutes rather than days.The derivation of the expression construct for the generation of the FRB expressing HeLa cell line is described in Motley (2006, PMID: 17035630)The Mito-YFP-FRB construct was based on a pEYFP-FRB plasmid (O. Glebov, MRC Laboratory of Molecular Biology, Cambridge, UK). The N-terminal sorting signal of Tom70p was amplified by PCR from yeast genomic DNA and cloned into the plasmid, then the Mito-YFP-FRB coding sequence was moved into the retroviral vector pQCXIH (Clontech), which carries a hygromycin resistance gene. Stable cell lines were selected using hygromyin selection.

General Info



Release Conditions

Restricted - commercial organisations are required to complete the 'Cell Line Release Authorisation for Research Use in Commercial Organisations' release conditions form in the supporting documents section.


Tissue of Origin





Study of protein function by rapid protein inactivation



Culture Conditions

Cell Type


Subculture Routine

Trypsinise cultures at 80% confluence with 0.05% Trypsin/EDTA. Seed flasks at 2x10⁴ cells/ cm². Cultures must be incubated in a humidified 10% CO₂/90% air incubator at 37°C.

Culture Medium

Dulbecco's Modified Eagle's Medium, high glucose (DMEM) + 2mM Glutamine + 10% Foetal Bovine Serum (FBS)

Growth Mode


Additional Info


Margaret S Robinson, University of Cambridge, Cambridge Institute for Medical Research, Cambridge CB2 0XY, UK

Country of Origin

United Kingdom

GMO Status

Genetically Modified Organism Class 1 (GMO1)

Hazard Group (ACDP)

Hazard Group (ACDP) 2



Robinson MS, Sahlender DA, Foster SD. Rapid inactivation of proteins by rapamycin-induced rerouting to mitochondria. Dev Cell. 2010 Feb 16;18(2):324-31. doi: 10.1016/j.devcel.2009.12.015. PMCID: PMC2845799, PMID: 20159602Hirst J, Borner GH, Antrobus R, Peden AA, Hodson NA, Sahlender DA, Robinson MS. Distinct and overlapping roles for AP-1 and GGAs revealed by the "knocksideways" system. Curr Biol. 2012 Sep 25;22(18):1711-6. doi: 10.1016/j.cub.2012.07.012. Epub 2012 Aug 16. PMCID: PMC3485558

Available Formats

  • Frozen
  • DNA-5µg (100ng/µl)

If use of this culture results in a scientific publication, it should be cited in the publication as: HeLa-Mitotrap (ECACC 15042201).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.