Product Class: Other

Sce PUS1
neb31 cloned at NEB recombinant 30 55 Heat

We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.


Catalog #M0526

Product Introduction

  • Sce PUS1 isomerizes uridine to pseudouridine in predominantly single-stranded RNA in vitro
  • Sce PUS1 is a stand-alone enzyme and does not require an RNA guide or co-factor to modify RNA
  • Sequence-specific pseudouridine modification by Sce PUS1 is an alternative to randomly incorporated modified nucleosides by RNA polymerases

This is an Enzyme for Innovation (EFI). EFI is a project initiated by NEB to provide unique enzymes to the scientific community in the hopes of enabling the discovery of new and innovative applications. These enzymes have interesting properties and unique specificities.

 

Product Information

Description

Sce Pseudouridine Synthase I (Sce PUS1) converts Uridine to Pseudouridine in single-stranded RNA, with a preference for Uridines in single-stranded RNA regions over Uridines in double-stranded RNA. The optimal substrate is an unstructured RNA that is 15 nt long or longer.

This product is related to the following categories:
Epitranscriptome Analysis,
RNA Modification,

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0526S     -20    
      Sce PUS1 M0526SVIAL -20 1 x 0.05 ml 100 pmol/μl
      NEBuffer™ r1.1 B6001SVIAL -20 1 x 1.25 ml 10 X

Properties & Usage

Reaction Conditions

1X NEBuffer™ r1.1
Incubate at 30°C

1X NEBuffer™ r1.1
10 mM Bis-Tris-Propane-HCl
10 mM MgCl2
100 µg/ml Recombinant Albumin
(pH 7 @ 25°C)

Heat Inactivation

55°C

References

  1. Arluison, V., Buckle, M., & Grosjean, H. (1999). Pseudouridine synthetase Pus1 of Saccharomyces cerevisiae: kinetic characterisation, tRNA structural requirement and real-time analysis of its complex with tRNA.  J Mol Bio. 289 (3), 491-502.
  2. Arluison, V., Batelier, G., Riès-Kautt, M., & Grosjean, H. (1999). RNA:pseudouridine synthetase Pus1 from Saccharomyces cerevisiae: oligomerization property and stoichiometry of the complex with yeast tRNA (Phe). Biochimie. 81 (7), 751-756.
  3. Carlile, T. M., Martinez, N. M., Schaening, C., Su, A., Bell, T. A., Zinshteyn, B., & Gilbert, W. V (2019). mRNA structure determines modification by pseudouridine synthase 1. Nature Chemical Biolog. 15 (10), 966-974.
  4. (2014). Pseudouridine profiling reveals regulated mRNA pseudouridylation in yeast and human cells. Nature. 515 (725), 143-146.
  5. (1998). The yeast tRNA:pseudouridine synthase Pus1p displays a multisite substrate specificity.. 4 (7), 856-869.
  6. Grünberg S, Doyle LA, Wolf EJ, Dai N, Corrêa IR Jr, Yigit E, et al (2023). The structural basis of mRNA recognition and binding by yeast pseudouridine synthase PUS1. 18(11), DOI: e0291267, PLoS ONE.

Protocols, Manuals & Usage

Protocols

  1. Protocol for conversion of Uridine to Pseudouridine using Sce PUS1 (NEB #M0526)

FAQs & Troubleshooting

FAQs

  1. What is the effect of Mg2+ concentration on Sce PUS1 activity?
  2. How can Sce PUS1 activity be improved when low concentrations of RNA are used?
  3. What effect does MnCl2 have on Sce PUS1 activity?
  4. What is the effect of NaCl concentration on Sce PUS1 activity?
  5. Can Sce PUS1 modify more than 1 µg of RNA in one reaction?
  6. Can I scale up or scale down the reaction volume of the Sce PUS1 reaction?
  7. Can I shorten or extend the incubation time of Sce PUS1 with target RNA?
  8. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

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.

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 busdev@neb.com.

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.