Improve Research Reproducibility A Bio-protocol resource
bpdetail_icon_lock
Preprint

Cell lines

ZZ Zuodong Zhao
ZZ Zhuqiang Zhang
BZ Bing Zhu

Last updated date: May 20, 2021 Views: 781 Forks: 0

original An abbreviated version of this protocol was published in eLife in Nov, 2020
Sustained TNF-α stimulation leads to transcriptional memory that greatly enhances signal sensitivity and robustness
download pdf Download PDF
ask Ask a question
cite How to cite
nfavorite Favorite

  The gRNAs we used are designed in silico via the CRISPR Design Tool (http://tools.genome-engineering.org). This CRISPR Design Tool is described in a Nature Protocol paper published Dr. Feng Zhang's group in 2013. The title of the Nature protocol paper is Genome engineering using the CRISPR-Cas9 system.

  2 days after the CRISPR plasmids transfection, we will treat the cells with puromycin for 1 day to select the transfected cells. Then we will seed the single cell into about 5 96-plates. When the clonies grow up, we will pick the edited clonies by genotyping.  

Copyright: Content may be subjected to copyright.
How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Zhao, Z, Zhang, Z and Zhu, B(2021). Cell lines. Bio-protocol Preprint. bio-protocol.org/prep1099.
  2. Zhao, Z., Zhang, Z., Li, J., Dong, Q., Xiong, J., Li, Y., Lan, M., Li, G. and Zhu, B.(2020). Sustained TNF-α stimulation leads to transcriptional memory that greatly enhances signal sensitivity and robustness. eLife. DOI: 10.7554/eLife.61965

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

0/150

tip Tips for asking effective questions

+ Description

Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.

post Post a Question
0 Q&A
This protocol preprint was submitted via the "Request a Protocol" track.