GAPTrap: A Simple Expression System for Pluripotent Stem Cells and Their Derivatives

Tim Kao, Tanya Labonne, Jonathan C. Niclis, Ritu Chaurasia, Zerina Lokmic, Elizabeth Qian, Freya F. Bruveris, Sara E Howden, Ali Motazedian, Jacqueline V. Schiesser, Magdaline Costa, Koula Sourris, Elizabeth Ng, David Anderson, Antonietta Giudice, Peter Farlie, Michael M H Cheung, Shireen R. Lamande, Anthony J Penington, Clare L. Parish & 5 others Lachlan H. Thomson, Arash Rafii, David A. Elliott, Andrew G. Elefanty, Edouard G. Stanley

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The ability to reliably express fluorescent reporters or other genes of interest is important for using human pluripotent stem cells (hPSCs) as a platform for investigating cell fates and gene function. We describe a simple expression system, designated GAPTrap (GT), in which reporter genes, including GFP, mCherry, mTagBFP2, luc2, Gluc, and lacZ are inserted into the GAPDH locus in hPSCs. Independent clones harboring variations of the GT vectors expressed remarkably consistent levels of the reporter gene. Differentiation experiments showed that reporter expression was reliably maintained in hematopoietic cells, cardiac mesoderm, definitive endoderm, and ventral midbrain dopaminergic neurons. Similarly, analysis of teratomas derived from GT-lacZ hPSCs showed that β-galactosidase expression was maintained in a spectrum of cell types representing derivatives of the three germ layers. Thus, the GAPTrap vectors represent a robust and straightforward tagging system that enables indelible labeling of PSCs and their differentiated derivatives.

Original languageEnglish
Pages (from-to)518-526
Number of pages9
JournalStem Cell Reports
Volume7
Issue number3
DOIs
Publication statusPublished - 13 Sep 2016

Keywords

  • differentiation
  • expression system
  • GAPDH
  • human pluripotent stem cells
  • lineage tracing
  • reporter genes

Cite this

Kao, Tim ; Labonne, Tanya ; Niclis, Jonathan C. ; Chaurasia, Ritu ; Lokmic, Zerina ; Qian, Elizabeth ; Bruveris, Freya F. ; Howden, Sara E ; Motazedian, Ali ; Schiesser, Jacqueline V. ; Costa, Magdaline ; Sourris, Koula ; Ng, Elizabeth ; Anderson, David ; Giudice, Antonietta ; Farlie, Peter ; Cheung, Michael M H ; Lamande, Shireen R. ; Penington, Anthony J ; Parish, Clare L. ; Thomson, Lachlan H. ; Rafii, Arash ; Elliott, David A. ; Elefanty, Andrew G. ; Stanley, Edouard G. / GAPTrap : A Simple Expression System for Pluripotent Stem Cells and Their Derivatives. In: Stem Cell Reports. 2016 ; Vol. 7, No. 3. pp. 518-526.
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Kao, T, Labonne, T, Niclis, JC, Chaurasia, R, Lokmic, Z, Qian, E, Bruveris, FF, Howden, SE, Motazedian, A, Schiesser, JV, Costa, M, Sourris, K, Ng, E, Anderson, D, Giudice, A, Farlie, P, Cheung, MMH, Lamande, SR, Penington, AJ, Parish, CL, Thomson, LH, Rafii, A, Elliott, DA, Elefanty, AG & Stanley, EG 2016, 'GAPTrap: A Simple Expression System for Pluripotent Stem Cells and Their Derivatives' Stem Cell Reports, vol. 7, no. 3, pp. 518-526. https://doi.org/10.1016/j.stemcr.2016.07.015

GAPTrap : A Simple Expression System for Pluripotent Stem Cells and Their Derivatives. / Kao, Tim; Labonne, Tanya; Niclis, Jonathan C.; Chaurasia, Ritu; Lokmic, Zerina; Qian, Elizabeth; Bruveris, Freya F.; Howden, Sara E; Motazedian, Ali; Schiesser, Jacqueline V.; Costa, Magdaline; Sourris, Koula; Ng, Elizabeth; Anderson, David; Giudice, Antonietta; Farlie, Peter; Cheung, Michael M H; Lamande, Shireen R.; Penington, Anthony J; Parish, Clare L.; Thomson, Lachlan H.; Rafii, Arash; Elliott, David A.; Elefanty, Andrew G.; Stanley, Edouard G.

In: Stem Cell Reports, Vol. 7, No. 3, 13.09.2016, p. 518-526.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Kao, Tim

AU - Labonne, Tanya

AU - Niclis, Jonathan C.

AU - Chaurasia, Ritu

AU - Lokmic, Zerina

AU - Qian, Elizabeth

AU - Bruveris, Freya F.

AU - Howden, Sara E

AU - Motazedian, Ali

AU - Schiesser, Jacqueline V.

AU - Costa, Magdaline

AU - Sourris, Koula

AU - Ng, Elizabeth

AU - Anderson, David

AU - Giudice, Antonietta

AU - Farlie, Peter

AU - Cheung, Michael M H

AU - Lamande, Shireen R.

AU - Penington, Anthony J

AU - Parish, Clare L.

AU - Thomson, Lachlan H.

AU - Rafii, Arash

AU - Elliott, David A.

AU - Elefanty, Andrew G.

AU - Stanley, Edouard G.

PY - 2016/9/13

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N2 - The ability to reliably express fluorescent reporters or other genes of interest is important for using human pluripotent stem cells (hPSCs) as a platform for investigating cell fates and gene function. We describe a simple expression system, designated GAPTrap (GT), in which reporter genes, including GFP, mCherry, mTagBFP2, luc2, Gluc, and lacZ are inserted into the GAPDH locus in hPSCs. Independent clones harboring variations of the GT vectors expressed remarkably consistent levels of the reporter gene. Differentiation experiments showed that reporter expression was reliably maintained in hematopoietic cells, cardiac mesoderm, definitive endoderm, and ventral midbrain dopaminergic neurons. Similarly, analysis of teratomas derived from GT-lacZ hPSCs showed that β-galactosidase expression was maintained in a spectrum of cell types representing derivatives of the three germ layers. Thus, the GAPTrap vectors represent a robust and straightforward tagging system that enables indelible labeling of PSCs and their differentiated derivatives.

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KW - expression system

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KW - lineage tracing

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