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A competitive precision CRISPR method to identify the fitness effects of transcription factor binding sites.

Published version
Peer-reviewed

Change log

Authors

Kauko, Otto 

Abstract

Here we describe a competitive genome editing method that measures the effect of mutations on molecular functions, based on precision CRISPR editing using template libraries with either the original or altered sequence, and a sequence tag, enabling direct comparison between original and mutated cells. Using the example of the MYC oncogene, we identify important transcriptional targets and show that E-box mutations at MYC target gene promoters reduce cellular fitness.

Description

Funder: Syöpäjärjestöt (Cancer Society of Finland); doi: https://doi.org/10.13039/501100006383


Funder: Jane ja Aatos Erkon Säätiö (Jane and Aatos Erkko Foundation); doi: https://doi.org/10.13039/501100004012


Funder: Sigrid Juséliuksen Säätiö (Sigrid Jusélius Foundation); doi: https://doi.org/10.13039/501100006306

Keywords

Binding Sites, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Cas Systems, Gene Editing, Promoter Regions, Genetic, Transcription Factors

Journal Title

Nat Biotechnol

Conference Name

Journal ISSN

1087-0156
1546-1696

Volume Title

41

Publisher

Springer Science and Business Media LLC
Sponsorship
Academy of Finland (Suomen Akatemia) (288836, 274555, 317807, 250345, 312042)
RCUK | Medical Research Council (MRC) (MR/V000500/1)
Cancer Research UK (CRUK) (RG99643)