CRISPR/Cas9 KOs of p53 and STAT1 in HEK293 cells

Haan, Tabitha, de (2022) CRISPR/Cas9 KOs of p53 and STAT1 in HEK293 cells. Bachelor's Research Project (period 2b), Biology.

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Abstract

Chromosomal instability is a well-known characteristic of cancer and leads to activation of inflammatory signalling. P53 and STAT1 are genes that play a role in this inflammatory response, while OAS1 and IFIT1 may also be involved. To better understand the underlying molecular mechanism, sgRNAs for these four genes were designed and cloned in vectors, to create CRISPR/Cas9 mediated knockout cells. HEK293 cells were transfected with vectors containing sgRNAs for p53 and STAT1. Western blot showed that STAT1 protein level was not decreased in cells targeted for STAT1 KO. The p53 protein level was slightly reduced in p53 KO targeted cells, while PCR and Sanger sequencing of the p53 gene in these cells showed no mutations in the sgRNA target sequence. We conclude that the CRISPR/Cas9 knockout of p53 and STAT1 in HEK293 cells was presumably unsuccessful.

Item Type:	Thesis (Bachelor's Research Project (period 2b))
Supervisor name:	Broekhuis, M.J.C.
Degree programme:	Biology
Thesis type:	Bachelor's Research Project (period 2b)
Language:	English
Date Deposited:	09 Jun 2022 09:49
Last Modified:	09 Jun 2022 09:49
URI:	https://fse.studenttheses.ub.rug.nl/id/eprint/27218

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