Toolkit/single cell-based analysis

single cell-based analysis

Computational Method·Research·Since 2019

Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.

Summary

Single cell-based analysis is a quantitative cellular assay framework developed to compare the activities of overexpressed full-length guanine nucleotide exchange factors in primary human endothelial cells. It was applied with single-cell FRET Rho GTPase biosensors to measure GEF-driven activation of Cdc42 and Rac1.

Usefulness & Problems

Why this is useful

This framework enables quantitative comparison of GEF efficiency at single-cell resolution in a primary endothelial cell context. In the cited study, it supported ranking endothelial GEFs for their ability to activate Cdc42 and assessing selectivity relative to Rac1.

Problem solved

The method addresses the problem of comparing the cellular activities of multiple overexpressed full-length GEFs in a quantitative manner. It also helps identify endothelial GEFs that may directly or indirectly activate Cdc42 using single-cell biosensor readouts.

Taxonomy & Function

Primary hierarchy

Technique Branch

Method: A concrete computational method used to design, rank, or analyze an engineered system.

Target processes

No target processes tagged yet.

Implementation Constraints

The reported implementation used single-cell FRET measurements with Rho GTPase biosensors in primary human endothelial cells. The assay compared overexpressed full-length GEF constructs, but the supplied evidence does not specify biosensor design, fluorophores, imaging hardware, or analysis pipeline details.

The supplied evidence describes application in one published study in primary human endothelial cells and does not establish performance across other cell types or signaling systems. The evidence also does not report throughput, sensitivity limits, statistical robustness, or independent replication.

Validation

Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication

Supporting Sources

Ranked Claims

Claim 1activity rankingsupports2019Source 1needs review

PLEKHG2, FGD1, PLEKHG1, and PREX1 induced the most efficient Cdc42 activation among the tested endothelial GEFs, and FGD1 showed the highest selectivity.

Our data reveal GEF dependent activation of Cdc42, with the most efficient Cdc42 activation induced by PLEKHG2, FGD1, PLEKHG1 and PREX1 and the highest selectivity for FGD1.
Claim 2activity rankingsupports2019Source 1needs review

PLEKHG2, FGD1, PLEKHG1, and PREX1 induced the most efficient Cdc42 activation among the tested endothelial GEFs, and FGD1 showed the highest selectivity.

Our data reveal GEF dependent activation of Cdc42, with the most efficient Cdc42 activation induced by PLEKHG2, FGD1, PLEKHG1 and PREX1 and the highest selectivity for FGD1.
Claim 3activity rankingsupports2019Source 1needs review

PLEKHG2, FGD1, PLEKHG1, and PREX1 induced the most efficient Cdc42 activation among the tested endothelial GEFs, and FGD1 showed the highest selectivity.

Our data reveal GEF dependent activation of Cdc42, with the most efficient Cdc42 activation induced by PLEKHG2, FGD1, PLEKHG1 and PREX1 and the highest selectivity for FGD1.
Claim 4activity rankingsupports2019Source 1needs review

PLEKHG2, FGD1, PLEKHG1, and PREX1 induced the most efficient Cdc42 activation among the tested endothelial GEFs, and FGD1 showed the highest selectivity.

Our data reveal GEF dependent activation of Cdc42, with the most efficient Cdc42 activation induced by PLEKHG2, FGD1, PLEKHG1 and PREX1 and the highest selectivity for FGD1.
Claim 5activity rankingsupports2019Source 1needs review

PLEKHG2, FGD1, PLEKHG1, and PREX1 induced the most efficient Cdc42 activation among the tested endothelial GEFs, and FGD1 showed the highest selectivity.

Our data reveal GEF dependent activation of Cdc42, with the most efficient Cdc42 activation induced by PLEKHG2, FGD1, PLEKHG1 and PREX1 and the highest selectivity for FGD1.
Claim 6activity rankingsupports2019Source 1needs review

PLEKHG2, FGD1, PLEKHG1, and PREX1 induced the most efficient Cdc42 activation among the tested endothelial GEFs, and FGD1 showed the highest selectivity.

Our data reveal GEF dependent activation of Cdc42, with the most efficient Cdc42 activation induced by PLEKHG2, FGD1, PLEKHG1 and PREX1 and the highest selectivity for FGD1.
Claim 7activity rankingsupports2019Source 1needs review

PLEKHG2, FGD1, PLEKHG1, and PREX1 induced the most efficient Cdc42 activation among the tested endothelial GEFs, and FGD1 showed the highest selectivity.

Our data reveal GEF dependent activation of Cdc42, with the most efficient Cdc42 activation induced by PLEKHG2, FGD1, PLEKHG1 and PREX1 and the highest selectivity for FGD1.
Claim 8assay applicationsupports2019Source 1needs review

Single cell FRET measurements with Rho GTPase biosensors were used in primary human endothelial cells to monitor GEF efficiency toward Cdc42 and Rac1.

By performing single cell FRET measurements with Rho GTPase biosensors in primary human ECs, we monitored GEF efficiency towards Cdc42 and Rac1.
Claim 9assay applicationsupports2019Source 1needs review

Single cell FRET measurements with Rho GTPase biosensors were used in primary human endothelial cells to monitor GEF efficiency toward Cdc42 and Rac1.

By performing single cell FRET measurements with Rho GTPase biosensors in primary human ECs, we monitored GEF efficiency towards Cdc42 and Rac1.
Claim 10assay applicationsupports2019Source 1needs review

Single cell FRET measurements with Rho GTPase biosensors were used in primary human endothelial cells to monitor GEF efficiency toward Cdc42 and Rac1.

By performing single cell FRET measurements with Rho GTPase biosensors in primary human ECs, we monitored GEF efficiency towards Cdc42 and Rac1.
Claim 11assay applicationsupports2019Source 1needs review

Single cell FRET measurements with Rho GTPase biosensors were used in primary human endothelial cells to monitor GEF efficiency toward Cdc42 and Rac1.

By performing single cell FRET measurements with Rho GTPase biosensors in primary human ECs, we monitored GEF efficiency towards Cdc42 and Rac1.
Claim 12assay applicationsupports2019Source 1needs review

Single cell FRET measurements with Rho GTPase biosensors were used in primary human endothelial cells to monitor GEF efficiency toward Cdc42 and Rac1.

By performing single cell FRET measurements with Rho GTPase biosensors in primary human ECs, we monitored GEF efficiency towards Cdc42 and Rac1.
Claim 13assay applicationsupports2019Source 1needs review

Single cell FRET measurements with Rho GTPase biosensors were used in primary human endothelial cells to monitor GEF efficiency toward Cdc42 and Rac1.

By performing single cell FRET measurements with Rho GTPase biosensors in primary human ECs, we monitored GEF efficiency towards Cdc42 and Rac1.
Claim 14assay applicationsupports2019Source 1needs review

Single cell FRET measurements with Rho GTPase biosensors were used in primary human endothelial cells to monitor GEF efficiency toward Cdc42 and Rac1.

By performing single cell FRET measurements with Rho GTPase biosensors in primary human ECs, we monitored GEF efficiency towards Cdc42 and Rac1.
Claim 15characterization summarysupports2019Source 1needs review

The study characterized endothelial GEFs that may directly or indirectly activate Cdc42.

Together, our study characterized endothelial GEFs that may directly or indirectly activate Cdc42
Claim 16characterization summarysupports2019Source 1needs review

The study characterized endothelial GEFs that may directly or indirectly activate Cdc42.

Together, our study characterized endothelial GEFs that may directly or indirectly activate Cdc42
Claim 17characterization summarysupports2019Source 1needs review

The study characterized endothelial GEFs that may directly or indirectly activate Cdc42.

Together, our study characterized endothelial GEFs that may directly or indirectly activate Cdc42
Claim 18characterization summarysupports2019Source 1needs review

The study characterized endothelial GEFs that may directly or indirectly activate Cdc42.

Together, our study characterized endothelial GEFs that may directly or indirectly activate Cdc42
Claim 19characterization summarysupports2019Source 1needs review

The study characterized endothelial GEFs that may directly or indirectly activate Cdc42.

Together, our study characterized endothelial GEFs that may directly or indirectly activate Cdc42
Claim 20characterization summarysupports2019Source 1needs review

The study characterized endothelial GEFs that may directly or indirectly activate Cdc42.

Together, our study characterized endothelial GEFs that may directly or indirectly activate Cdc42
Claim 21characterization summarysupports2019Source 1needs review

The study characterized endothelial GEFs that may directly or indirectly activate Cdc42.

Together, our study characterized endothelial GEFs that may directly or indirectly activate Cdc42
Claim 22method developmentsupports2019Source 1needs review

A new single cell-based analysis was developed to enable quantitative comparison of cellular activities of overexpressed full-length GEFs.

A new, single cell-based analysis was developed and used to enable the quantitative comparison of cellular activities of the overexpressed full-length GEFs.
Claim 23method developmentsupports2019Source 1needs review

A new single cell-based analysis was developed to enable quantitative comparison of cellular activities of overexpressed full-length GEFs.

A new, single cell-based analysis was developed and used to enable the quantitative comparison of cellular activities of the overexpressed full-length GEFs.
Claim 24method developmentsupports2019Source 1needs review

A new single cell-based analysis was developed to enable quantitative comparison of cellular activities of overexpressed full-length GEFs.

A new, single cell-based analysis was developed and used to enable the quantitative comparison of cellular activities of the overexpressed full-length GEFs.
Claim 25method developmentsupports2019Source 1needs review

A new single cell-based analysis was developed to enable quantitative comparison of cellular activities of overexpressed full-length GEFs.

A new, single cell-based analysis was developed and used to enable the quantitative comparison of cellular activities of the overexpressed full-length GEFs.
Claim 26method developmentsupports2019Source 1needs review

A new single cell-based analysis was developed to enable quantitative comparison of cellular activities of overexpressed full-length GEFs.

A new, single cell-based analysis was developed and used to enable the quantitative comparison of cellular activities of the overexpressed full-length GEFs.
Claim 27method developmentsupports2019Source 1needs review

A new single cell-based analysis was developed to enable quantitative comparison of cellular activities of overexpressed full-length GEFs.

A new, single cell-based analysis was developed and used to enable the quantitative comparison of cellular activities of the overexpressed full-length GEFs.
Claim 28method developmentsupports2019Source 1needs review

A new single cell-based analysis was developed to enable quantitative comparison of cellular activities of overexpressed full-length GEFs.

A new, single cell-based analysis was developed and used to enable the quantitative comparison of cellular activities of the overexpressed full-length GEFs.

Approval Evidence

1 source1 linked approval claimfirst-pass slug single-cell-based-analysis
A new, single cell-based analysis was developed and used to enable the quantitative comparison of cellular activities of the overexpressed full-length GEFs.

Source:

method developmentsupports

A new single cell-based analysis was developed to enable quantitative comparison of cellular activities of overexpressed full-length GEFs.

A new, single cell-based analysis was developed and used to enable the quantitative comparison of cellular activities of the overexpressed full-length GEFs.

Source:

Comparisons

Source-backed strengths

The assay was specifically developed for quantitative comparison of full-length GEF activities at the single-cell level. In primary human endothelial cells, it resolved differential Cdc42 activation efficiency, identifying PLEKHG2, FGD1, PLEKHG1, and PREX1 as the strongest activators among those tested, with FGD1 showing the highest selectivity.

Ranked Citations

  1. 1.
    StructuralSource 1Small GTPases2019Claim 1Claim 2Claim 3

    Extracted from this source document.