Evidence map›Paper›PMID 39936976›Full record

ArticleCells2025

A Recombinase-Mediated Cassette Exchange Platform for a Triple Independent Inducible Expression System for Human Pluripotent Stem Cells.

Roberto Castro-Gutierrez, Ankita Arora, Katherine F Vaeth, J Matthew Taliaferro, Holger A Russ

Abstract read
In one paragraph

Article in Cells, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

5 authors.

Roberto Castro-GutierrezDiabetes Institute, College of Medicine, University of Florida, Gainesville, FL 32610, USA.ORCID 0000-0001-5873-6473
Ankita AroraDepartment of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.
Katherine F VaethDepartment of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.
J Matthew TaliaferroDepartment of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.ORCID 0000-0001-7580-1433
Holger A RussDiabetes Institute, College of Medicine, University of Florida, Gainesville, FL 32610, USA.ORCID 0000-0001-5117-2927

Funding

Renewal of the Human Islet Research Enhancement Center (HIREC) for the Type-1-Diabetes-Focused Human Islet Research Network (HIRN).U24DK104162 · NIDDK · BECKMAN RESEARCH INSTITUTE/CITY OF HOPE · PI John S. Kaddis, Joyce Carol Niland · 2019 to 2026
$12.3M
University of Colorado Anschutz Medical Campus DRCP30DK116073 · NIDDK · UNIVERSITY OF COLORADO DENVER · PI LORI SUSSEL · 2020 to 2026
$10.8M
Understanding the regulatory language of RNA localizationR35GM133385 · NIGMS · UNIVERSITY OF COLORADO DENVER · PI Jefferson Matthew Taliaferro · 2019 to 2026
$3.4M
Improving human pluripotent stem cell derived beta cell transplantation using genetic lineage tracing.R01DK120444 · NIDDK · UNIVERSITY OF COLORADO DENVER · PI RUSS, HOLGER A. · 2018 to 2021
$1.5M
Deciphering the nanomaterial uptake in a bystander manner invoked by co-administration of functionalized nanoparticlesR01GM133885 · NIGMS · UNIVERSITY OF MINNESOTA · PI PANG, HONGBO · 2019 to 2022
$1.2M
Stress induced changes to the human beta cell proteomeR21AI140044 · NIAID · UNIVERSITY OF COLORADO DENVER · PI DAVIDSON, HOWARD W, RUSS, HOLGER A. · 2019 to 2020
$428k
NIAID NIH HHS R21 AI140044NIDDK NIH HHS P30 DK116073NIDDK NIH HHS R01 DK120444NIDDK NIH HHS U24 DK104162NIGMS NIH HHS R01 GM133885NIGMS NIH HHS R35 GM133385NIGMS NIH HHS R35 GM133885
6 · The paper itself

Abstract

Human pluripotent stem cells (hPSCs) and their differentiated derivatives represent valuable tools for studying development, modeling diseases, and advancing cell therapy. Recent improvements in genome engineering allow for precise modifications of hPSCs, further enhancing their utility in basic and translational research. Here we describe a Recombinase-Mediated Cassette Exchange (RMCE) platform in hPSCs that allows for the highly efficient, rapid, and specific integration of transgenes. The RCME-mediated DNA integration process is nearly 100% efficient, without negatively affecting the pluripotency or karyotypic stability of hPSCs. Taking advantage of this convenient system, we first established a dual inducible expression system based on the Tet-On and Cumate-On systems, allowing for the inducible expression of two transgenes independently. Secondly, we incorporated a Tet-on inducible system, driving the expression of three genes simultaneously. However, two genes also contain independent degron sequences, allowing for precise control over the expression of each gene individually. We demonstrated the utility of these systems in hPSCs, as well as their functionality after differentiation into cells that were representative of the three germ layers. Lastly, we used the triple inducible system to investigate the lineage commitment induced by the pancreatic transcription factors NKX6.1 and PDX1. We found that controlled dual expression, but not individual expression, biases hPSC embryoid body differentiation towards the pancreatic lineage by inducing the expression of the NeuroD program. In sum, we describe a novel genetic engineering platform that allows for the efficient and fast integration of any desired transgene(s) in hPSCs using RMCE. We anticipate that the ability to modulate the expression of three transgenes simultaneously will further accelerate discoveries using stem cell technology.

Indexed as

Pluripotent Stem CellsRecombinasesCell DifferentiationHumansTransgenesRecombinasescumatedegrondual expression systemgenome engineeringgerm layer differentiationhuman pluripotent stem cellsNKX6.1pancreaticPDX1recombinase-mediated cassette exchange (RMCE)tetracyclinetranscription factorstriple expression system

Identifiers

PMID39936976
PMCPMC11817695

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.