Next Article in Journal
CD28 Autonomous Signaling Up-Regulates C-Myc Expression and Promotes Glycolysis Enabling Inflammatory T Cell Responses in Multiple Sclerosis
Next Article in Special Issue
S. cerevisiae Strain Lacking Mitochondrial IF3 Shows Increased Levels of Tma19p during Adaptation to Respiratory Growth
Previous Article in Journal
MiR-501-3p Forms a Feedback Loop with FOS, MDFI, and MyoD to Regulate C2C12 Myogenesis
Previous Article in Special Issue
Regulation of Ribosomal Proteins on Viral Infection
Open AccessArticle

A Versatile Strategy to Reduce UGA-Selenocysteine Recoding Efficiency of the Ribosome Using CRISPR-Cas9-Viral-Like-Particles Targeting Selenocysteine-tRNA[Ser]Sec Gene

1
Centre International de Recherche en Infectiologie (CIRI), 69007 Lyon, France
2
Institut National de la Santé et de la Recherche Médicale (INSERM) Unité U1111, 69007 Lyon, France
3
Ecole Normale Supérieure de Lyon, 69342 Lyon, France
4
Université Claude Bernard Lyon 1 (UCBL1), 69622 Lyon, France
5
Unité Mixte de Recherche 5308 (UMR5308), Centre National de la Recherche Scientifique (CNRS), 69007 Lyon, France
*
Authors to whom correspondence should be addressed.
Cells 2019, 8(6), 574; https://doi.org/10.3390/cells8060574
Received: 28 May 2019 / Revised: 7 June 2019 / Accepted: 10 June 2019 / Published: 11 June 2019
(This article belongs to the Special Issue Ribosome Function and Dynamics)
The translation of selenoprotein mRNAs involves a non-canonical ribosomal event in which an in-frame UGA is recoded as a selenocysteine (Sec) codon instead of being read as a stop codon. The recoding machinery is centered around two dedicated RNA components: The selenocysteine insertion sequence (SECIS) located in the 3′ UTR of the mRNA and the selenocysteine-tRNA (Sec-tRNA[Ser]Sec). This translational UGA-selenocysteine recoding event by the ribosome is a limiting stage of selenoprotein expression. Its efficiency is controlled by the SECIS, the Sec-tRNA[Ser]Sec and their interacting protein partners. In the present work, we used a recently developed CRISPR strategy based on murine leukemia virus-like particles (VLPs) loaded with Cas9-sgRNA ribonucleoproteins to inactivate the Sec-tRNA[Ser]Sec gene in human cell lines. We showed that these CRISPR-Cas9-VLPs were able to induce efficient genome-editing in Hek293, HepG2, HaCaT, HAP1, HeLa, and LNCaP cell lines and this caused a robust reduction of selenoprotein expression. The alteration of selenoprotein expression was the direct consequence of lower levels of Sec-tRNA[Ser]Sec and thus a decrease in translational recoding efficiency of the ribosome. This novel strategy opens many possibilities to study the impact of selenoprotein deficiency in hard-to-transfect cells, since these CRISPR-Cas9-VLPs have a wide tropism. View Full-Text
Keywords: selenium; selenocysteine; selenoprotein; SECIS; Sec-tRNA[Ser]Sec; UGA-recoding; CRISPR-Cas9; viral-like particles; nanoblades selenium; selenocysteine; selenoprotein; SECIS; Sec-tRNA[Ser]Sec; UGA-recoding; CRISPR-Cas9; viral-like particles; nanoblades
Show Figures

Graphical abstract

MDPI and ACS Style

Vindry, C.; Guillin, O.; Mangeot, P.E.; Ohlmann, T.; Chavatte, L. A Versatile Strategy to Reduce UGA-Selenocysteine Recoding Efficiency of the Ribosome Using CRISPR-Cas9-Viral-Like-Particles Targeting Selenocysteine-tRNA[Ser]Sec Gene. Cells 2019, 8, 574.

Show more citation formats Show less citations formats
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