Product Class: Other

RNase R
NEBU recombinant engineered 37

Product Introduction

  • Highly processive 3´ to 5´ exoribonuclease
  • Digests nearly all linear RNAs with an accessible 3´ end, including rRNA and mRNA
  • Enriches for circular RNAs and lariat RNAs from total RNA preps
  • Requires magnesium for activity
  • Inactivated with the addition of excess EDTA
Catalog # Size Concentration
M0100S 400 units 20000 units/ml

Product Information

Description

Ribonuclease R (RNase R) is a processive 3´ to 5´ exoribonuclease (1,2). RNase R requires a 3´ single-stranded RNA substrate that is approximately 10 nucleotides or longer, such as a poly (A) tail, for proper binding (3,4,5). RNase R is unique from other exoribonucleases for its ability to degrade highly structured RNAs without the need for an additional helicase (2,6,7). Circular RNAs and lariat RNAs are enriched after RNase R digestion as these closed RNA structures are resistant to exoribonucleases (8).


Figure 1. RNase R digests linear RNA with an accessible 3´ end

Mechanism of RNase R, showing digestion of linear RNA and tail of lariat RNA, but no access to circular RNA

RNase R digests linear RNA with an accessible 3´ end and can be used to enrich for circular and lariat RNA. RNase R can be used to digest linear RNAs. Circular RNAs are closed RNA molecules that are resistant to RNase R digestion. Lariat RNAs have a looped structure with a single-stranded RNA region at the 3´ end. RNase R can digest the linear RNA at the 3´ end, but stops at the branch point of the lariat, preserving the looped RNA.


Figure 2. RNase R efficiently degrades linear RNA and leaves circular RNA intact

Gels showing optimal performance of RNase R for linear RNA, circular RNA and both

NEB® RNase R efficiently degrades linear RNA and leaves circular RNA intact. Adding 1 unit of NEB RNase R per 1 µg of RNA sample containing only linear RNA, only circular RNA, or both linear and circular RNA results in circular RNA maintenance and linear RNA digestion. This reaction ran for 15 or 30 minutes at 37°C in 1X RNase R Reaction Buffer.



 

Product Source

An E. coli strain that carries the cloned RNase R gene (rnr) from Escherichia coli with an N-terminal 6xHis-tag.
This product is related to the following categories:
RNases,
RNA Modification,

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0100S     -20    
      RNase R M0100SVIAL -20 1 x 0.02 ml 20,000 units/ml
      RNase R Reaction Buffer B0100SVIAL -20 2 x 1.25 ml 10 X

Properties & Usage

Materials Required but not Supplied

Unit Definition

One unit is defined as the amount of enzyme required to convert 75 pmoles of 20-nucleotide single-stranded RNA sequence downstream of a 38-nucleotide DNA hairpin into acid soluble ribonucleotides in a total reaction volume of 20 µl in 15 minutes at 25°C.

Reaction Conditions

1X RNase R Reaction Buffer
Incubate at 37°C

1X RNase R Reaction Buffer
20 mM Tris-HCl
100 mM KCl
0.1 mM MgCl2

Storage Buffer

300 mM NaCl
10 mM Tris-HCl
0.1 mM EDTA
1 mM DTT
surfactant
50% Glycerol
pH 7.4 @ 25°C

References

  1. Kasai T, et al. (1977). Exoribonucleases in wild type Escherichia coli and RNase II-deficient mutants. J Biol Chem. Dec 25; 252 (24), 8950-6. PubMedID: 336626
  2. Cheng ZF and Deutscher MP. (2002). Purification and characterization of the Escherichia coli exoribonuclease RNase R. J Biol Chem. Jun 14; 277 (24), 21624-9. PubMedID: 11948193
  3. Cheng ZF and Deutscher MP. (2005). An important role for RNase R in mRNA decay. Mol Cell. Jan 21; 17(2), 313-8. PubMedID: 15664199
  4. Vincent HA and Deutscher MP. (2006). Substrate recognition and catalysis by the exoribonuclease RNase R. J Biol Chem. Oct 6;281(40), 29769-75. PubMedID: 16893880
  5. Xiao MS and Wilusz JE. (2019). An improved method for circular RNA purification using RNase R that efficiently removes linear RNAs containing G-quadruplexes or structured 3' ends. Nucleic Acids Res. Sep 19; 47 (16), 8755-69. PubMedID: 31269210
  6. Hossain ST, et al. (2016). How RNase R Degrades Structured RNA: ROLE OF THE HELICASE ACTIVITY AND THE S1 DOMAIN. J Biol Chem. Apr 8; 291 (15), 7877-87. PubMedID: 26872969
  7. Awano N, et al. (2010). Escherichia coli RNase R has dual activities, helicase and RNase. J. Bacteriol. Mar; 192 (5), 1344-52. PubMedID: 20023028
  8. Suzuki H, et al. (2006). Characterization of RNase R-digested cellular RNA source that consists of lariat and circular RNAs from pre-mRNA splicing. Nucleic Acids Res. May 8; 34 (8), e63. PubMedID: 16682442

Protocols, Manuals & Usage

Protocols

  1. Digestion of Linear RNA with RNase R to Enrich for Circular RNA (NEB #M0100)

FAQs & Troubleshooting

FAQs

  1. Is RNase R (NEB #M0100) active at temperatures other than 37°C?
  2. Is it necessary to clean up RNA samples before RNase R (NEB #M0100) digestion?
  3. How long does the 3´ single-stranded RNA overhang need to be for RNase R (NEB #M0100) to bind?
  4. Can RNase R (NEB #M0100) be heat-inactivated?
  5. What are circular RNAs?
  6. Does RNase R (NEB #M0100) have star activity?
  7. Is it possible to digest RNase R (NEB #M0100)-resistant linear RNAs?
  8. Can RNase R diluted in 1X Reaction Buffer be stored at 4°C?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email [email protected].

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.