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Blood Center of Wisconsin

2 ARTICLES PUBLISHED IN JoVE

Cancer Research

Investigation of Genetic Dependencies Using CRISPR-Cas9-based Competition Assays

Anagha Deshpande¹, Bo Rui Chen¹, Luyi Zhao¹, Kelsey Saddoris¹, Mayuri Kerr¹, Nan Zhu², Prashant Mali³, Aniruddha J. Deshpande¹

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 Barnekow¹, Jordan Palmer², Adam Beck³, 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.