Poly(dA:dT) Rhodamine

Rhodamine labeled double-stranded B-DNA

Poly(dA:dT) Rhodamine was chemically labeled by covalent coupling of a rhodamine probe containing a reactive alkylating group. This confers fluorescent properties to Poly(dA:dT) with a slight reduction of pattern recognition receptor (PRR) stimulatory activity.

Poly(dA:dT) is a poly(deoxyadenylic-deoxythymidylic) acid sodium salt and a repetitive double-stranded DNA (dsDNA) sequence of poly(dA-dT)•poly(dT-dA). Poly(dA:dT) is a synthetic analog of B-DNA.

Intracellular poly(dA:dT) is recognized by several cytosolic DNA sensors, triggering an immune response.

Furthermore, poly(dA:dT) is indirectly sensed by RIG-I leading to type I IFN production.

Specifications

Activity: RIG-I & CDS agonist, AIM2 inflammasome inducer

CAS number: 86828-69-5

Endotoxin level: Spectral properties of rhodamine:
Excitation λ max: 546 nm
Emission λ max: 576 nm

Applications:
Poly(dA:dT) Rhodamine can be used for various applications:

• flow cytometry
• fluorescent and confocal microscopy

Contents

Poly(dA:dT) Rhodamine is provided lyophilized.

• 10 mg Poly(dA:dT) Rhodamine
• 1.5 ml endotoxin-free water

Poly(dA:dT) Rhodamine is shipped at room temperature.

Description

Poly(dA:dT) Rhodamine was chemically labeled by covalent coupling of a rhodamine probe containing a reactive alkylating group. This confers fluorescent properties to Poly(dA:dT) with a slight reduction of pattern recognition receptor (PRR) stimulatory activity. Poly(dA:dT) is a poly(deoxyadenylic-deoxythymidylic) acid sodium salt and a repetitive double-stranded DNA (dsDNA) sequence of poly(dA-dT)•poly(dT-dA). Poly(dA:dT) is a synthetic analog of B-DNA.
Intracellular poly(dA:dT) is recognized by several cytosolic DNA sensors, such as ZBP1/DAI and LRRFIP1, triggering an immune response [1-5]. ZBP1/DAI and LRRFIP1 bind poly(dA:dT) inducing type I IFN production via TBK1/IRF-3 and β-catenin pathways, respectively [3,4].
Additionally, poly(dA:dT) is recognized by AIM2 triggering the formation of an inflammasome and the subsequent secretion of IL-1β and IL-18 [5]. Furthermore, transfected poly(dA:dT) can be transcribed by RNA polymerase III into dsRNA with a 5’-triphosphate moiety (5’ppp-dsRNA) which is a ligand for RIG-I [6,7]. Thus poly(dA:dT) is indirectly sensed by RIG-I leading to type I IFN production through the adaptor molecule IPS-1 and the TBK1/IRF3 pathway [8].

References:

1. Ishii KJ, et al., 2006. A Toll-like receptor-independent antiviral response induced by double-stranded B-form DNA. Nat Immunol. 7(1):40-8.
2. Ishii KJ. & akira S., 2006. Innate immune recognition of, and regulation by, DNA Trends Immunol. 27(11):525-32.
3. Takaoka a. et al., 2007. DAI (DLM-1/ZBP1) is a cytosolic DNA sensor and an activator of innate immune response. Nature. 448(7152):501-5.
4. Yang P. et al., 2010. The cytosolic nucleic acid sensor LRRFIP1 mediates the production of type I interferon via a beta-catenin-dependent pathway. Nat Immunol. 11(6):487-94.
5. Jones JW. et al., 2010. Absent in melanoma 2 is required for innate immune recognition of Francisella tularensis. PNAS, 107(21):9771-6.
6. Ablasser a. et al., 2009. RIG-I-dependent sensing of poly(dA:dT) through the induction of an RNA polymerase III-transcribed RNA intermediate. Nat Immunol. 10(10):1065-72.
7. Chiu YH. et al., 2009. RNA polymerase III detects cytosolic DNA and induces type I interferons through the RIG-I pathway. Cell. 138(3):576-91.
8. Takeshita f. & ishii KJ., 2008. Intracellular DNA sensors in immunity. Curr Opin Immunol. 20(4):383-8.