Dihxy cells, developed from DEN-injected C57BL/6 mice, were generously provided by Dr. Michael Karin’s laboratory and cultured as previously described (6 (link)). The cells were mycoplasma free as detected by Mycoplasma Detection Kit (ThermoFisher) and were not used for more than 10 passages. These cells were transfected with a plasmid expressing either control, scrambled sgRNA or AEG-1 sgRNA, Cas9, puromycin-resistance marker and mCherry (obtained from GeneCopoeia). Single clones from FACS-sorted mCherry positive cells were isolated, expanded and validated for AEG-1 knockout.
Protocol detail
Generation and Validation of AEG-1 Knockout Cells
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : Virginia Commonwealth University Medical Center
Other organizations : The University of Texas Health Science Center at San Antonio
Top 5 similar protocols
Variable analysis
independent variables
- Transfection with control, scrambled sgRNA or AEG-1 sgRNA, Cas9, puromycin-resistance marker and mCherry
- AEG-1 knockout
- Dihxy cells developed from DEN-injected C57BL/6 mice
- Cells were mycoplasma free
- Cells were not used for more than 10 passages
- Positive control: Cells transfected with control or scrambled sgRNA
- Negative control: Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!