Blood Center of Wisconsin
2 ARTICLES PUBLISHED IN JoVE
Cancer Research
Investigation of Genetic Dependencies Using CRISPR-Cas9-based Competition Assays
Anagha Deshpande 1, Bo Rui Chen 1, Luyi Zhao 1, Kelsey Saddoris 1, Mayuri Kerr 1, Nan Zhu 2, Prashant Mali 3, Aniruddha J. Deshpande 1
1Tumor Initiation and Maintenance Program, Sanford Burnham Prebys Medical Discovery Institute, 2Blood Research Institute, Blood Center of Wisconsin, 3Department of Bioengineering, University of California, San Diego
This manuscript describes a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) CRISPR-Cas9-based method for simple and expeditious investigation of the role of multiple candidate genes in Acute Myeloid Leukemia (AML) cell proliferation in parallel. This technique is scalable and can be applied in other cancer cell lines as well.
Biochemistry
Translating Ribosome Affinity Purification (TRAP) for RNA Isolation from Endothelial Cells In Vivo
Patrick Moran 1,2, Yichen Guo 3,4, Rong Yuan 1,3, Nicholas Barnekow 1, Jordan Palmer 2, Adam Beck 3, Bin Ren 3,4,5
1Blood Research Institute, Blood Center of Wisconsin, 2Department of Medicine, Medical College of Wisconsin, 3Department of Surgery, The University of Alabama at Birmingham, 4Department of Biomedical Engineering, The University of Alabama at Birmingham, 5GBS Program, Graduate School, The University of Alabama at Birmingham
We present an approach to purify ribosome-bound mRNA from vascular endothelial cells (ECs) directly in mouse brain, lung and heart tissues via EC-specific genetic tag of enhanced green fluorescence protein (EGFP)in ribosomes in combination with RNA purification.
