Mammalian Reverse Genetics without Crossing Reveals Nr3a as a Short-Sleeper Gene

Cell Reports
Genshiro A SunagawaHiroki R Ueda

Abstract

The identification of molecular networks at the system level in mammals is accelerated by next-generation mammalian genetics without crossing, which requires both the efficient production of whole-body biallelic knockout (KO) mice in a single generation and high-performance phenotype analyses. Here, we show that the triple targeting of a single gene using the CRISPR/Cas9 system achieves almost perfect KO efficiency (96%-100%). In addition, we developed a respiration-based fully automated non-invasive sleep phenotyping system, the Snappy Sleep Stager (SSS), for high-performance (95.3% accuracy) sleep/wake staging. Using the triple-target CRISPR and SSS in tandem, we reliably obtained sleep/wake phenotypes, even in double-KO mice. By using this system to comprehensively analyze all of the N-methyl-D-aspartate (NMDA) receptor family members, we found Nr3a as a short-sleeper gene, which is verified by an independent set of triple-target CRISPR. These results demonstrate the application of mammalian reverse genetics without crossing to organism-level systems biology in sleep research.

References

Mar 2, 2002·Science·Hiroaki Kitano
Nov 15, 2002·Nature·Hiroaki Kitano
Mar 20, 2004·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Priyattam J ShiromaniDavid R Weaver
Apr 29, 2005·Nature·Chiara CirelliGiulio Tononi
Jul 19, 2005·Nature Neuroscience·Eric M SnyderPaul Greengard
Feb 7, 2007·IEEE Transactions on Bio-medical Engineering·Aaron E FloresBruce F O'Hara
Sep 18, 2007·Respiratory Research·Thomas GlaabWayne Mitzner
Jan 9, 2010·Science·Philippe Horvath, Rodolphe Barrangou
Jan 26, 2010·Progress in Neurobiology·Maile A HensonBenjamin D Philpot
Jul 16, 2010·Genetics·Iara D CarberyXiaoxia Cui
Oct 1, 2011·Neuropsychopharmacology : Official Publication of the American College of Neuropsychopharmacology·Bita Moghaddam, Daniel Javitt
Feb 18, 2012·Nature·Blake WiedenheftJennifer A Doudna
Mar 2, 2012·Sleep·Blakeley B McShaneAllan I Pack
Jan 11, 2013·Nature Biotechnology·Young Hoon SungHan-Woong Lee
Apr 30, 2013·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Genshiro A SunagawaHiroki R Ueda
Sep 3, 2013·Nucleic Acids Research·Wataru FujiiKunihiko Naito
Oct 26, 2013·Nature Protocols·F Ann RanFeng Zhang
Jan 28, 2014·Nature Biotechnology·Yanfang FuJ Keith Joung
Feb 6, 2014·The FEBS Journal·Jiankui ZhouXingxu Huang

❮ Previous
Next ❯

Citations

May 24, 2016·Journal of Genetics and Genomics = Yi Chuan Xue Bao·Maximilian Haeussler, Jean-Paul Concordet
Jan 27, 2017·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Kohei HommaMakoto Kaneda
May 20, 2017·Expert Opinion on Drug Discovery·Mitsuyuki MatsumotoKatsunori Tajinda
Dec 1, 2017·Nature Protocols·Hideki UkaiHiroki R Ueda
Jun 5, 2019·Proceedings. Biological Sciences·S NealE C Seaver
Nov 14, 2019·Journal of Cell Science·Katsumi FumotoAkira Kikuchi
Oct 2, 2019·Proceedings of the National Academy of Sciences of the United States of America·Shogo MatobaAtsuo Ogura
Feb 22, 2017·Frontiers in Neurology·Arthur Millius, Hiroki R Ueda
Aug 2, 2017·Mammalian Genome : Official Journal of the International Mammalian Genome Society·Lydia TeboulPatrick M Nolan
Sep 19, 2018·Proceedings of the National Academy of Sciences of the United States of America·Kensuke YoshidaHiroki R Ueda
Mar 14, 2019·Nucleic Acids Research·Bo ZhouAlexander E Urban
Jun 13, 2020·Nature·Tohru M TakahashiTakeshi Sakurai
Jun 27, 2017·NPJ Systems Biology and Applications·Etsuo A SusakiHiroki R Ueda
Feb 6, 2020·Frontiers in Neuroscience·Rikuhiro G Yamada, Hiroki R Ueda
Feb 12, 2020·Journal of Personalized Medicine·Chiara ArgentatiSabata Martino
Feb 10, 2018·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Tetsushi SakumaTakashi Yamamoto
Dec 6, 2017·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Shoi Shi, Hiroki R Ueda
Jul 4, 2018·Scientific Reports·Toru SuzukiAnthony C F Perry
Apr 11, 2020·F1000Research·Maxime JanPaul Franken
Dec 26, 2019·BMC Genomics·Jacob Bradford, Dimitri Perrin
Aug 30, 2019·PLoS Computational Biology·Jacob Bradford, Dimitri Perrin
Nov 11, 2020·Proceedings of the National Academy of Sciences of the United States of America·Phillip A ClevesJohn R Pringle
Dec 9, 2020·Cerebral Cortex·Alvaro MurilloIsabel Pérez-Otaño
Oct 31, 2020·Frontiers in Psychology·Koji L Ode, Hiroki R Ueda
Feb 5, 2021·Scientific Reports·Alexandra LahtinenTiina Paunio
Apr 8, 2019·Current Opinion in Biotechnology·Hideki UkaiHiroki R Ueda
Aug 30, 2018·Cell Reports·Yasutaka NiwaHiroki R Ueda
Mar 29, 2021·Nanomedicine : Nanotechnology, Biology, and Medicine·Keisuke UmemuraMakiya Nishikawa
Apr 24, 2017·Neuroscience Research·Fumiya TatsukiHiroki R Ueda
Dec 30, 2018·Neuroscience Research·Asako NoguchiYuji Ikegaya
May 13, 2021·Journal of Radiation Research·Kento MiuraOsamu Kaminuma
May 5, 2021·The Journal of Physiology·Oliver CrawleyIsabel Pérez-Otaño
Jul 23, 2021·Current Biology : CB·Hiroshi KiyonariYasuhide Furuta
Dec 29, 2020·Current Opinion in Neurobiology·John M Webb, Ying-Hui Fu
Dec 14, 2021·Frontiers in Cell and Developmental Biology·Nasir Javaid, Sangdun Choi

❮ Previous
Next ❯

Related Concepts

Related Feeds

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.

Cell Imaging in CNS

Here is the latest research on cell imaging and imaging modalities, including light-sheet microscopy, in the central nervous system.

Related Papers

Molecular Therapy : the Journal of the American Society of Gene Therapy
Dana Carroll
Journal of Sleep Research
Derk-Jan Dijk
Annals of the New York Academy of Sciences
Helena T Mueller, James H Meador-Woodruff
© 2022 Meta ULC. All rights reserved