Cite as: Cold Spring Harb. Protoc.; 2009; doi:10.1101/pdb.prot5270

This Protocol
Right arrow Full Text
Right arrow Update/discuss this protocolDiscussion icon
Right arrow Alert me when this protocol is cited
Right arrow Alert me when comments are published
Right arrow Alert me if a correction is posted
Services
Right arrow Similar protocols in this database
Right arrow Similar articles in PubMed
Right arrow Alert me to new releases of protocols
Right arrow Save to Personal Folders
Right arrow Download to citation manager
Right arrow Printer-friendly versionPrinter-friendly version
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Davidson, A. E.
Right arrow Articles by Krull, C. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Davidson, A. E.
Right arrow Articles by Krull, C. E.
Related Collections
Right arrow Cell Biology, general
Right arrow Cell Culture
Right arrow Molecular Biology, general
Right arrow Analysis of Gene Expression
Right arrow Transgenic Technology, general
Right arrow Transgenic Mice
Right arrow Transgenic Mice, general
Right arrow Developmental Biology
Right arrow DNA Delivery/Gene Transfer
Right arrow Stem Cells
Right arrow Embryonic Stem Cells
Right arrow DNA Delivery/Gene Transfer, general
Right arrow Non-Viral Methods
Right arrow Analysis of Gene Expression in Cultured Cells
Right arrowRelated Protocols
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

protocolProtocol

Use of the Sleeping Beauty Transposon System for Stable Gene Expression in Mouse Embryonic Stem Cells

Ann E. Davidson1, Theresa E. Gratsch2, Maria H. Morell2, K. Sue O’Shea2, and Catherine E. Krull1,3

1 Department of Biologic and Materials Sciences, University of Michigan, Ann Arbor, MI 48109, USA
2 Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA

3Corresponding author (krullc{at}umich.edu)


INTRODUCTION

Sleeping Beauty (SB) transposon-based transfection is a two-component system consisting of a transposase and a transposon containing inverted repeat/direct repeat (IR/DR) sequences that result in precise integration into a TA dinucleotide. The transposon is designed with an expression cassette of interest flanked by IR/DRs, and SB transposase mediates stable integration and reliable long-term expression of the gene of interest. It has recently been demonstrated that SB efficiently mediates gene transfer and stable gene expression in human embryonic stem (ES) cells. Here, we describe a method for transfecting and establishing stable cell lines in mouse embryonic stem (mES) cells with the SB system.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?

Related Protocols

Estimation of Cell Number by Hemocytometry Counting
Joseph Sambrook and David W. Russell
Cold Spring Harb Protoc 2006: 4454. [Extract] [Full Text]

Inverse PCR
Joseph Sambrook and David W. Russell
Cold Spring Harb Protoc 2006: 3487. [Extract] [Full Text]

Amplification of cDNA Generated by Reverse Transcription of mRNA
Joseph Sambrook and David W. Russell
Cold Spring Harb Protoc 2006: 3837. [Extract] [Full Text]

Southern Blotting: Capillary Transfer of DNA to Membranes
Joseph Sambrook and David W. Russell
Cold Spring Harb Protoc 2006: 4040. [Extract] [Full Text]

Southern Blotting: Simultaneous Transfer of DNA from a Single Agarose Gel to Two Membranes
Joseph Sambrook and David W. Russell
Cold Spring Harb Protoc 2006: 4043. [Extract] [Full Text]

Southern Hybridization of Radiolabeled Probes to Nucleic Acids Immobilized on Membranes
Joseph Sambrook and David W. Russell
Cold Spring Harb Protoc 2006: 4044. [Extract] [Full Text]