Next Article in Journal
Translational Prospects and Challenges in Human Induced Pluripotent Stem Cell Research in Drug Discovery
Next Article in Special Issue
Invariant Chain Complexes and Clusters as Platforms for MIF Signaling
Previous Article in Journal
Potential of Induced Pluripotent Stem Cells (iPSCs) for Treating Age-Related Macular Degeneration (AMD)
Previous Article in Special Issue
Role of Cytokine-Induced Glycosylation Changes in Regulating Cell Interactions and Cell Signaling in Inflammatory Diseases and Cancer
Article

Random Splicing of Several Exons Caused by a Single Base Change in the Target Exon of CRISPR/Cas9 Mediated Gene Knockout

Institute of Biochemistry, Medical Faculty, University of Giessen, Friedrichstrasse 24, D-35392 Giessen, Germany
*
Author to whom correspondence should be addressed.
Academic Editor: Alexander E. Kalyuzhny
Cells 2016, 5(4), 45; https://doi.org/10.3390/cells5040045
Received: 20 October 2016 / Revised: 3 December 2016 / Accepted: 9 December 2016 / Published: 14 December 2016
(This article belongs to the Special Issue Signal Transduction 2016)
The clustered regularly interspaced short palindromic repeats (CRISPR)-associated sequence 9 (CRISPR/Cas9) system is widely used for genome editing purposes as it facilitates an efficient knockout of a specific gene in, e.g. cultured cells. Targeted double-strand breaks are introduced to the target sequence of the guide RNAs, which activates the cellular DNA repair mechanism for non-homologous-end-joining, resulting in unprecise repair and introduction of small deletions or insertions. Due to this, sequence alterations in the coding region of the target gene frequently cause frame-shift mutations, facilitating degradation of the mRNA. We here show that such CRISPR/Cas9-mediated alterations in the target exon may also result in altered splicing of the respective pre-mRNA, most likely due to mutations of splice-regulatory sequences. Using the human FLOT-1 gene as an example, we demonstrate that such altered splicing products also give rise to aberrant protein products. These may potentially function as dominant-negative proteins and thus interfere with the interpretation of the data generated with these cell lines. Since most researchers only control the consequences of CRISPR knockout at genomic and protein level, our data should encourage to also check the alterations at the mRNA level. View Full-Text
Keywords: RNA splicing; CRISPR; Cas9; genome editing; flotillin RNA splicing; CRISPR; Cas9; genome editing; flotillin
Show Figures

Figure 1

MDPI and ACS Style

Kapahnke, M.; Banning, A.; Tikkanen, R. Random Splicing of Several Exons Caused by a Single Base Change in the Target Exon of CRISPR/Cas9 Mediated Gene Knockout. Cells 2016, 5, 45. https://doi.org/10.3390/cells5040045

AMA Style

Kapahnke M, Banning A, Tikkanen R. Random Splicing of Several Exons Caused by a Single Base Change in the Target Exon of CRISPR/Cas9 Mediated Gene Knockout. Cells. 2016; 5(4):45. https://doi.org/10.3390/cells5040045

Chicago/Turabian Style

Kapahnke, Marcel, Antje Banning, and Ritva Tikkanen. 2016. "Random Splicing of Several Exons Caused by a Single Base Change in the Target Exon of CRISPR/Cas9 Mediated Gene Knockout" Cells 5, no. 4: 45. https://doi.org/10.3390/cells5040045

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop