Evidence map›Paper›PMID 32474864›Full record

ArticleMethods in molecular biology (Clifton, N.J.)2020

Generation of an Induced Pluripotent Stem Cell Line with the Constitutive EGFP Reporter.

Kiel T Butterfield, Patrick S McGrath, Chann Makara Han, Igor Kogut, Ganna Bilousova

Abstract read
In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing 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

2 citing papers in PubMed.

  1. Review
  2. CIRCLE-Seq for Interrogation of Off-Target Gene Editing.Journal of visualized experiments : JoVE · 2024
    Article
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.

Kiel T ButterfieldDepartment of Dermatology, University of Colorado School of Medicine, Aurora, CO, USA.
Patrick S McGrathDepartment of Dermatology, University of Colorado School of Medicine, Aurora, CO, USA.
Chann Makara HanDepartment of Dermatology, University of Colorado School of Medicine, Aurora, CO, USA.
Igor KogutDepartment of Dermatology, University of Colorado School of Medicine, Aurora, CO, USA.
Ganna BilousovaDepartment of Dermatology, University of Colorado School of Medicine, Aurora, CO, USA. ganna.bilousova@cuanschutz.edu.

Funding

Transgenic and Gene Targeting CoreP30AR057212 · NIAMS · UNIVERSITY OF COLORADO DENVER · PI NORRIS, DAVID A. · 2009 to 2018
$6.2M
MicroRNA-based purification of keratinocytes derived from pluripotent stem cells for the treatment of skin diseasesR21AR074642 · NIAMS · UNIVERSITY OF COLORADO DENVER · PI BILOUSOVA, GANNA · 2019 to 2020
$376k
NIAMS NIH HHS P30 AR057212NIAMS NIH HHS R21 AR074642
6 · The paper itself

Abstract

The discovery of induced pluripotent stem cell (iPSC) technology has provided a versatile platform for basic science research and regenerative medicine. With the rise of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) systems and the ease at which they can be utilized for gene editing, creating genetically modified iPSCs has never been more advantageous for studying both organism development and potential clinical applications. However, to better understand the behavior and true therapeutic potential of iPSCs and iPSC-derived cells, a tool for labeling and monitoring these cells in vitro and in vivo is needed. Here, we describe a protocol that provides a straightforward method for introducing a stable, highly expressed fluorescent protein into iPSCs using the CRISPR/Cas9 system and a standardized donor vector. The approach involves the integration of the EGFP transgene into the transcriptionally active adeno-associated virus integration site 1 (AAVS1) locus through homology directed repair. The knockin of this transgene results in the generation of iPSC lines with constitutive expression of the EGFP protein that also persists in differentiated iPSCs. These EGFP-labeled iPSC lines are ideal for assessing iPSC differentiation in vitro and evaluating the distribution of iPSC-derived cells in vivo after transplantation into model animals.

Indexed as

Gene ExpressionGenes, ReporterGenetic EngineeringCell Culture TechniquesCell DifferentiationCells, CulturedCRISPR-Cas SystemsGene EditingGene Knock-In TechniquesGene TargetingGenetic VectorsGreen Fluorescent ProteinsHumansInduced Pluripotent Stem Cellsenhanced green fluorescent proteinGreen Fluorescent ProteinsCRISPR/Cas9Fluorescent proteinInduced pluripotent stem cellsiPSCsNucleofection

Identifiers

PMID32474864
PMCPMC7396154

What OpenQuestion holds

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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.