Name: Drew Kyla L.

Baysa
Date: August 24, 2019

Title of Journal Article: CRISPR/Cas9 as a tool to dissect cancer mutations

Authors: Shady Sayed, Maciej Paszkowski-Rogacz, Lukas Theo Schmitt, Frank Buchholz
Date published: 10 May 2019

Summary:

Studies about CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/ Cas9
(CRISPR-associated protein 9) technology has been rampant nowadays. It emerged as a very
powerful technology for genome editing and is being utilized in the fields of biotechnology,
genetics, microbiology and medicine. Specifically in the field of medicine, it has been used to
correct disease causing DNA mutations ranging from a single base pair to large deletions in model
systems ranging from cells in vitro to animals in vivo. However, not all mutations contribute equally
to turmorigenesis and being able to distinguish specific mutations for tumor growth. This paper
presents a method to screen for the functional relevance of mutations in high throughput in
established cancer cell lines. It utilizes the CRISPR/Cas9 system to probe cancer vulnerabilities
in a colorectal carcinoma cell line in an attempt to identify novel cancer driver mutations. The
analysis revealed two sgRNAs targeting the same mutation (UTP14A: S99delS) to be depleted
over time in RKO cells. Validation and characterization confirmed that the inactivation of this
mutation impairs cell growth, nominating UTP14A: S99delS as a putative driver mutation in RKO
cells. Overall, it demonstrates that the CRISPR/Cas9 system is a powerful tool to functionally
dissect.