We believe this site might serve you best:

United States

United States

Choose language: English

Promega's Cookie Policy

Our website uses functional cookies that do not collect any personal information or track your browsing activity. When you select your country, you agree that we can place these functional cookies on your device.

A Practical Guide to CRISPR-Mediated Gene Tagging with a Bioluminescent Peptide

Dr. Schwinn is a member of the cell biology team within the Advanced Technologies Group at Promega where she has helped develop luciferase-based technologies for studying intracellular protein interactions, abundance, and post-translational modifications.  Most recently, she has focused on combining these technologies with CRISPR/Cas9 to study protein dynamics at the endogenous level.  Prior to joining Promega, Dr. Schwinn earned her doctorate in Biochemistry from the University of Wisconsin-Madison in the lab of Dr. Hector F. DeLuca, and she received post-doctoral training from Dr. Donna M. Peters in the Department of Pathology and Laboratory Medicine.
  • Marie Schwinn, PhD

  • Sr Research Scientist
    Promega Corporation

Tuesday, October 24, 2017

CRISPR/Cas9 gene editing has emerged as a powerful tool for making precise genomic modifications. Although often used to create gene knockouts, the technology also enables targeted knock-in of a specific sequence. An exciting application is the endogenous tagging of proteins expressed under native regulatory conditions. In this webinar, we will detail a simple and efficient cloning-free method for CRISPR-mediated endogenous gene tagging, focusing on knock-in of the sensitive, bioluminescent HiBiT tag.

Webinar Summary:

CRISPR-Cas9 technology creates double-strand breaks (DSBs) at almost any genomic location of interest. Following the DSB, cellular repair machinery can either undergo non-homologous end joining (NHEJ) creating knockouts, or, if a donor DNA template is provided, the cell can undergo homology directed repair (HDR) to knock-in a precise modification or insertion. Often researchers may want to knock-in a tag sequence, enabling further study of protein biology under endogenous regulatory conditions. Compared to ectopic expression of a tagged protein, endogenous locus expression eliminates artifacts of overexpression, mislocalization and aberrant signaling to provide a better understanding of native protein biology. Many existing protein tags are large, which makes CRISPR-mediated knock-in inefficient. Antibody-based detection methods and low sensitivity make many protein tags undesirable for use as endogenous tags. We recently developed the HiBiT bioluminescent protein tagging system, which provides a tagging option that is amenable to CRISPR-mediated gene tagging. The HiBiT tagging system contains a tag of only 11 amino acids with great sensitivity rendering simple antibody-free protein detection. In this webinar, we will demonstrate an efficient method for CRISPR-mediated HiBiT tagging that requires no molecular cloning steps so that you can progress from gene editing to assaying endogenous biology in as little as 24 hours.

Share With a Friend

We will NOT sell your personal information, such as your name or email address, to third parties or use your email address to send you marketing materials. The email and names entered will only be used to identify the sender and recipient of the email.

Send Cancel

Time Zone Converter

Date

Time

Convert To

Converted Time

Choose your country

Americas

Brazil
Canada
United States

Pacific Asia

Australia
India
Japan
Korea, Republic of
Singapore

Europe

Austria
Belgium
Denmark
Estonia
Finland
France
Germany
Iceland
Italy
Luxembourg
Netherlands
Norway
Poland
Spain
Sweden
Switzerland
United Kingdom