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NATE™

NATE™ Unit size Cat. code Docs Qty Price
Nucleic Acid Transfection Enhancer
1 mL (100 reactions)
lyec-nate
+-
$128.00

NATE™ - Nucleic Acid Transfection Enhancer
NATE™ - Nucleic Acid Transfection Enhancer

NEW PRODUCT

Boost your transfection with NATE™

 

NATE™ is a nucleic acid transfection enhancer designed by InvivoGen to boost both transient and stable transfection efficiencies in hard-to-transfect cells; specifically human monocytes and murine macrophages (i.e. THP-1 and RAW 264.7 cells, respectively). Simply add NATE™ 30 mins prior to your current transfection protocol. 

The principal obstacle for ‘foreign’ nucleic acids (i.e. plasmids) during eukaryotic cell transfection is their own detection by cytosolic DNA sensors of the innate immune system (see 'Details' tab). These defensive cellular strategies can greatly affect the efficiency of both transient and stable transfections. When using NATE™, a number of these nucleic acid sensing pathways will be inhibited, thereby protecting the plasmid and facilitating its expression.

 

Key Features of NATE™:

  • Compatible with commonly used transfection reagents (e.g. GeneXPlus, Lipofectamine® LTX, and jetPRIME®) as well as physical methods (e.g. nucleofection).
  • Higher transfection yield, even with large plasmids (> 10kb).
  • Gentle on cells with no toxicity in all tested transfection protocols.
     

NATE™ is a highly pure (>95%) proprietary blend of innate immune system inhibitors and has been functionally tested in transfection assays using THP-1 and RAW 264.7 cells. 

 

Figures

Greater transient transfection efficiency in NATE™ treated THP-1 cells
Greater transient transfection efficiency in NATE™ treated THP-1 cells

Top - Transient transfection of an ~3 kb GFP-expressing plasmid into THP-1 cells was performed using GeneXPlus both in the absence (left) and presence (right) of NATE™. After 48 hours, the transfected cells expressing GFP were visualized by fluorescence microscopy. Bottom - Transient transfections of an ~3 kb GFP-expressing plasmid (left) and an 8 kb GFP-expressing plasmid (right) into THP-1 cells was performed using commonly used transfection methods such as GeneXPlus, Lipofectamine® LTX, jetPRIME®, and nucleofection. This was performed both in the presence (green) and absence (yellow) of NATE™. After 48 hours, transfection efficiency (% GFP-expressing cells) was measured using flow cytometry. Data are presented as a fold change compared to transfection without NATE™ 

Greater transient transfection efficiency in NATE™ treated RAW 264.7 cells
Greater transient transfection efficiency in NATE™ treated RAW 264.7 cells

Top - Transient transfection of an ~3 kb GFP-expressing plasmid into RAW 264.7 cells was performed using Lipofectamine® LTX both in the absence (left) and presence (right) of NATE™. After 48 hours, the transfected cells expressing GFP were visualized by fluorescence microscopy. Bottom - Transient transfection of an ~3 kb GFP-expressing plasmid (left) and an 8 kb GFP-expressing plasmid (right) into RAW 264.7 cells was performed using commonly used transfection methods including Lipofectamine® LTX, jetPRIME®, FuGENE®, and nucleofection. This was performed both in the presence (green) and absence (yellow) of NATE™. After 48 hours, transfection efficiency (% GFP-expressing cells) was measured using flow cytometry. Data are presented as fold change compared to transfection without NATE™.

Increased number of stable SEAP-expressing RAW 264.7 clones
Increased number of stable SEAP-expressing RAW 264.7 clones

Stable transfection of an ~10 kb SEAP‑expressing plasmid into RAW 264.7 cells was performed using Lipofectamine® LTX both in the absence (left) and presence (right) of NATE™. After 10 days in selection with Blasticidin, the number of stable clones expressing SEAP (blue wells) was visualized using QUANTI‑Blue™, a SEAP detection reagent.

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Product video & Details

 

 

 

Details

Nucleic acid sensing pathways encountered during transfection
Nucleic acid sensing pathways encountered during transfection

The principal obstacle for ‘foreign’ nucleic acids (such as plasmid DNA) during eukaryotic cell transfection is their own detection by cytosolic nucleic acid sensors such as Absent in melanoma 2 (AIM2) and cyclic GMP-AMP synthase (cGAS) [1].

AIM2 is a cytosolic sensor that strongly responds to double-stranded DNA (dsDNA) leading to the formation of a protein complex called the inflammasome. Ultimately, the activation of the AIM2 inflammasome induces the maturation of the pro-inflammatory cytokines’ interleukin-1β (IL-1β) and IL-18 via caspase-1 cleavage. The production of these cytokines is detrimental to transfection success [1].

Furthermore, the sensing of dsDNA by cGAS induces the production of type I interferons (IFNs), through the STING/TBK1/IRF3 signaling axis, as well as the expression of a number of interferon-stimulated genes (ISGs), which exert potent effector functions, which are highly unwanted in transfection [1].

Additionally, the activation of STING can lead to LC3-dependent autophagy, through a  TBK1- and IFN-independent mechanism [2]. Eventually, an autophagosome will form containing the 'foreign' nucleic acids (e.g. plasmid), and fuse with a lysosome for its clearance from the cell.

 

References:

1. Patrick, K.L. et al. 2016. For Better or Worse: Cytosolic DNA Sensing during Intracellular Bacterial Infection Induces Potent Innate Immune Responses. J Mol Biol 428, 3372-3386.
2. Gui, X. et al. 2019. Autophagy induction via STING trafficking is a primordial function of the cGAS pathway. Nature 567, 262-266.

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Specifications

Composition: Proprietary blend of innate immune system inhibitors

Purity: > 95% UHPLC

Quality Control:

  • Absence of bacterial contamination (i.e. lipoproteins and endotoxin) has been confirmed using HEK-Blue™  TLR2 and TLR4 cellular assays, respectively.
  • Biological activity of NATE™ has been confirmed using transfection assays.

 

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Contents

  • 2 vials of NATE™ (approximately 100 reactions) provided in an evaporated form

Room temperature Product is shipped at room temperature

Store Upon receipt product should be stored at -20°C

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