An all-in-one UniSam vector system for efficient gene activation

Antonella Fidanza; Martha Lopez yrigoyen; Nicola Romano; Rhiannon Jones; Alice Taylor; Lesley M. Forrester

doi:10.1038/s41598-017-06468-6

An all-in-one UniSam vector system for efficient gene activation

Antonella Fidanza, Martha Lopez yrigoyen, Nicola Romano, Rhiannon Jones, Alice Taylor, Lesley M. Forrester

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Abstract / Description of output

We have generated a drug-free, all-in-one dCAS9-SAM vector that can activate endogenous gene expression with the potential to modify cell fate. We demonstrate that this strategy can be used in a number of cell lines and avoids exceptionally high levels of gene expression that are observed in standard transgenic approaches. Compared to the multi-plasmid system, this all-in-one vector activates gene expression to a comparable level but the reduced overall DNA content results in significantly higher viability of transfected cells. This allowed us to use the RUNX1C-GFP human embryonic stem cell reporter cell line to monitor gene activation in individual cells and to show that activation could occur at all stages of the cell cycle.

Original language	English
Article number	6394
Journal	Scientific Reports
Early online date	25 Jul 2017
DOIs	https://doi.org/10.1038/s41598-017-06468-6
Publication status	E-pub ahead of print - 25 Jul 2017

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10.1038/s41598-017-06468-6

s41598-017-06468-6
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Lesley Forrester
- Deanery of Clinical Sciences - UoE Retired Staff
- Centre for Regenerative Medicine
- Centre for Inflammation Research
- Edinburgh Haematopoiesis Network
Person: Academic: Research Active , Affiliated Independent Researcher

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title = "An all-in-one UniSam vector system for efficient gene activation",

abstract = "We have generated a drug-free, all-in-one dCAS9-SAM vector that can activate endogenous gene expression with the potential to modify cell fate. We demonstrate that this strategy can be used in a number of cell lines and avoids exceptionally high levels of gene expression that are observed in standard transgenic approaches. Compared to the multi-plasmid system, this all-in-one vector activates gene expression to a comparable level but the reduced overall DNA content results in significantly higher viability of transfected cells. This allowed us to use the RUNX1C-GFP human embryonic stem cell reporter cell line to monitor gene activation in individual cells and to show that activation could occur at all stages of the cell cycle.",

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AU - Romano, Nicola

AU - Jones, Rhiannon

AU - Taylor, Alice

AU - Forrester, Lesley M.

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AB - We have generated a drug-free, all-in-one dCAS9-SAM vector that can activate endogenous gene expression with the potential to modify cell fate. We demonstrate that this strategy can be used in a number of cell lines and avoids exceptionally high levels of gene expression that are observed in standard transgenic approaches. Compared to the multi-plasmid system, this all-in-one vector activates gene expression to a comparable level but the reduced overall DNA content results in significantly higher viability of transfected cells. This allowed us to use the RUNX1C-GFP human embryonic stem cell reporter cell line to monitor gene activation in individual cells and to show that activation could occur at all stages of the cell cycle.

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An all-in-one UniSam vector system for efficient gene activation

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Lesley Forrester

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