Boost your transfections 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. 

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.

Contents

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

 Product is shipped at room temperature

 Upon receipt product should be stored at -20°C

Details

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.

如需购买或咨询该产品技术问题，请联系Invivogen一级代理商--欣博盛生物