Lentiviral Biosensors

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The GFP-LC3 displays a diffuse nuclear and cytosolic distribution in fed cells, and a punctate distribution in starved autophagic cells.
HeLa cells were transduced with LentiBrite™GFP-LC3 lentiviral particles and were either (A) left in complete media or (B) incubated in EBSS containing a lysosome inhibitor under starvation conditions. The GFP-LC3 displays a diffuse nuclear and cytosolic distribution in fed cells, and a punctate distribution in starved autophagic cells.
Biosensors can be used to detect a particular protein as well as the subcellular location of that protein within live cells. Fluorescent tags are an effective way to visualize the protein of interest within a cell by either fluorescent microscopy or time-lapse video capture. Visualizing live cells without disruption can reveal changing cellular conditions in real time.

Lentiviral vector systems are a popular tool for introducing genes and gene products into cells. Advantages over non-viral methods (such as chemical-based transfection) include higher-efficiency transfection of dividing and non-dividing cells, stable expression of the transgene, and low immunogenicity.

MilliporeSigma’s new LentiBrite™ Lentiviral Biosensors, a new suite of pre-packaged lentiviral particles encoding important and foundational proteins of autophagy, apoptosis, and cell structure, enables visualization under different cell/disease states in live cell and in vitro analysis.

Features and Benefits:
  • Pre-packaged, ready-to-use, fluorescently-tagged with monomeric GFP & RFP
  • Minimum titer (≥3 x 10^8 IFU/mL) per vial
  • Higher efficiency transfection as compared to traditional chemical-based and other non-viral-based transfection methods
  • Ability to transfect dividing, non-dividing, and difficult-to-transfect cell types, such as primary cells or stem cells
  • Non-disruptive towards cellular function
  • Validated for fluorescent microscopy and live cell analysis
Live Cell Visualization

LentiBrite GFP-LC3 Lentiviral Biosensor,
 Catalogue No. 17-10193

Timecourse imaging of HT-1080 cells after transduction with LentiBrite GFP-LC3 and the addition of a lysosomal inhibitor under starvation conditions. GFP-LC3 displays a diffuse nuclear and cytosolic distribution in fed cells, and a punctate distribution in starved, autophagic cells.




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LentiBrite EB3-GFP Lentiviral Biosensor,
 Catalogue No. 17-10208

REF52 cells were plated in a chambered coverglass and transduced with lentiviral particles. Cells were imaged live by oil immersion wide-field fluorescence microscopy. The EB3-GFP displays a punctate distribution characteristic of microtubule ends.




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LentiBrite RFP-EB3 Lentiviral Biosensor,
 Catalogue No. 17-10222

REF52 cells were plated in a chambered coverglass and transduced with lentiviral particles. Cells were imaged live by oil immersion wide-field fluorescence microscopy. The EB3-RFP displays a punctate distribution characteristic of microtubule ends.




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LentiBrite Histone H2B-RFP Lentiviral Biosensor,
 Catalogue No. 17-10228

U2OS cells were plated in chamber slides and transduced with lentiviral particles. Cells were imaged live by oil immersion wide-field fluorescence microscopy. The cell displays metaphase chromosomes (H2B-RFP) as it transitions to anaphase.




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LentiBrite Histone H2B-GFP Lentiviral Biosensor,
 Catalogue No. 17-10229

U2OS cells were plated in chamber slides and transduced with lentiviral particles. Cells were imaged live by oil immersion wide-field fluorescence microscopy. The cell is displaying prophase, where the chromatin (H2B-GFP) condenses into chromosomes.




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Higher Efficiency Transfection Compared to Chemical or Non-viral-based Transfection Methods
GFP-Tubulin (HUVEC)
HUVECs were transfected with GFP-Tubulin plasmids using Lipofectamine 2000 (top row) or LentiBrite GFP-Tubulin lentiviral particles (bottom row).   Lentibrite GFP-Tubulin HUVEC transfection using Lipofectamine 2000.
Plasmid Transfection Using Lipofectamine 2000
Tubulin HUVEC transfection using Lentibrite GFP-Tubulin.   GFP-Tubulin HUVEC transfection using Lentibrite GFP-Tubulin.
LentiBrite GFP-Tubulin Lentiviral Biosensor Tranfection
(Catalogue No. 17-10206)
HUVECs were transfected with GFP-Tubulin plasmids using Lipofectamine 2000 (top row) or LentiBrite GFP-Tubulin lentiviral particles (bottom row). The nucleus is stained with DAPI (blue). The LentiBrite lentiviral transfection yields higher efficiency and more uniform expression of GFP-Tubulin versus chemical transfection.

Higher Efficiency Transfection Compared to Baculovirus
MOI = 20

LentiBrite
LentiBrite lentiviral particles encoding GFP-LC3 were adjusted to 1 mL, and equivalent volumes of lentivirus and baculovirus from a competitor product were added to HT1080 cells at an MOI of 20.   LentiBrite lentiviral particles encoding GFP-LC3 were adjusted to 1 mL, and equivalent volumes of lentivirus and baculovirus from a competitor product were added to HT1080 cells at an MOI of 20.
Baculovirus Competitor Product
LentiBrite lentiviral particles encoding GFP-LC3 were adjusted to 1 mL, and equivalent volumes of lentivirus and baculovirus from a competitor product were added to HT1080 cells at an MOI of 20.   LentiBrite lentiviral particles encoding GFP-LC3 were adjusted to 1 mL, and equivalent volumes of lentivirus and baculovirus from a competitor product were added to HT1080 cells at an MOI of 20.
LentiBrite lentiviral particles encoding GFP-LC3 (Catalogue No. 17-10193) were adjusted to 1 mL, and equivalent volumes of lentivirus and baculovirus from a competitor product were added to HT1080 cells at an MOI of 20. Lentiviral transfection provides a stronger overall signal of GFP-LC3 compared to baculovirus transfection, when using equivalent amounts of each product.

Ability to Transfect Dividing, Non-dividing, and Difficult-to-transfect Cell Types
LentiBrite Paxillin-GFP primary cells.   LentiBrite RFP-Beta Actin primary cells.   LentiBrite GFP-Vimentin primary cells.
LentiBrite Paxillin-GFP (Catalogue No. 17-10154) Primary cells (Human Mesenchymal Stem Cells) were plated in chamber slides and transduced with lentiviral particles at an MOI of 5 for 24 hours.
 
LentiBrite RFP-Beta Actin (Catalogue No. 17-10203) Primary cells (HUVECs) were plated in chamber slides and transduced with lentiviral particles at an MOI of 40 for 24 hours.

 
LentiBrite GFP-Vimentin (Catalogue No. 17-10152) Primary cells (Human Mesenchymal Stem Cells) were plated in chamber slide and transduced with lentiviral particles at an MOI of 10 for 24 hours.

LentiBrite Lentiviral Biosensor Family
DescriptionCatalogue No.
GFP-LC3 17-10193
GFP-LC3 Mutant 17-10189
RFP-LC3 17-10143
RFP-LC3 Control Mutant 17-10188

Alpha-actinin-GFP 17-10156
Alpha-Actinin-RFP 17-10196

GFP-Beta Actin 17-10204
RFP-Beta Actin 17-10203

Calreticulin-RFP 17-10146

PSD95-GFP 17-10227
PSD95-RFP 17-10226

EB3-GFP 17-10208
EB3-RFP 17-10222
DescriptionCatalogue No.
Histone H2B-GFP 17-10229
Histone H2B-RFP 17-10228

GFP-HMGB1 17-10147

Paxillin-GFP 17-10154
Paxillin-RFP 17-10155

RFP-Tubulin 17-10205
GFP-Tubulin 17-10206

GFP-Vimentin 17-10152
RFP-Vimentin 17-10153

GFP-RAD51 17-10234
RFP-RAD51 17-10220

GFP-Cortactin 17-10219
RFP-Cortactin 17-10218

More GFP- & RFP-tagged lentiviral biosensors coming soon!