Reliable CRISPR/Cas9 genome engineering in Caenorhabditis elegans using a single efficient sgRNA and an easily recognizable phenotype

wrmScarlet is much brighter than TagRFP-T in vivo.

wrmScarlet is much brighter than TagRFP-T.
wrmScarlet is codon-optimized of the bright monomeric fluorescent protein mScarlet invented by the lab of Theodorus W J Gadella Jr (molecularcytology.nl).

We generated two CRISPR knockin lines that fuse either TagRFP-T or wrmScarlet to the N-terminus of the two-pore domain potassium channel TWK-18.

In this comparison, wrmScarlet was on average 8 times brighter than TagRFP-T.

Plasmids. They are available right now. Free. To anyone who needs them. #ASAPbio

To efficiently and freely distribute the wrmScarlet sequence to the community, we have decided to list here all the labs that we have sent the sequence to. Please contact us or anyone on the list to get your copy of the pSEM87 plasmid.

To design your own wrmScarlet clone, you can download the annotated sequence file here pSEM87. "Right-click" on the file name and use "Save to Link As..." to save the file on your hard drive. This annotated sequence file is in Genbank format and can be opened with most software packages. We like ApE and benchling.com.

  • (25) Europe
    • (6) France
      • Lyon - Jean-Louis Bessereau {INMG}
      • Lyon - Maïté Carré-Pierrat {UMS3421}
      • Strasbourg - Sophie Jarriault {IGBMC}
      • Bordeaux - Denis Dupuy {Institut Europeen de Chimie et Biologie}
      • Paris - Michel Labouesse {UPMC}
      • Paris - Marie Anne Félix {ENS Paris}
    • (1) Austria
      • Vienna - Luisa Cochella {IMP}
    • (6) Switzerland
      • Basel - Iskra Katic {Friedrich Miescher Institute for Biomedical Research}
      • Basel - Rafal Ciosk {Friedrich Miescher Institute for Biomedical Research} (via I. Katic)
      • Basel - Helge Grosshans {Friedrich Miescher Institute for Biomedical Research} (via I. Katic)
      • Basel - Susan M. Gasser {Friedrich Miescher Institute for Biomedical Research} (via I. Katic)
      • Bern - Peter Meister (via Iskra Katic) {University of Bern}
      • Geneva - Florian Steiner {University of Geneva}
    • (4) Netherlands
      • Utrecht - Mike Boxem >> Sander van den Heuvel, Casper Hoogenraad {Utrecht University}
      • Wageningen - Jan Kammenga {Wageningen University & Research}
    • (5) Germany
      • Freiburg - Stefan Eimer {BIOSS}
      • Berlin - Baris Tursun {Max Delbrueck Center for Molecular Medicine, Berlin}
      • Berlin - Nikolaus Rajewsky {Max Delbrueck Center for Molecular Medicine, Berlin}
      • Planegg - Adeline Mergoud (Hartl departement) {Max-Planck-Institute of Biochemistry, Martinsried}
      • Mainz - René Ketting {Institute for Molecular Biology (IMB) gGmbH}
    • (2) Spain
      • Sevilla - Peter Askjaer {Centro Andaluz de Biología del Desarrollo}
      • Barcelona - Julián Cerón Madrigal {Bellvitge Biomedical Research Institute (IDIBELL)}
    • (1) Polska
      • Warsaw - Michał Turek {Institute of Molecular and Cell Biology}
    • (1) Ireland
      • Dublin - Oliver Blacque {UCD Conway Institute of Biomolecular and Biomedical Research}

  • (1) Turkey
    • Kayseri - Sebiha Cevik-Kaplan {Abdullah Gul University}

  • (9) United Kingdom
    • London - Michalis Barkoulas {Imperial College}
    • London - Richard Poole {University College London}
    • London - Nathan Goehring {the Francis Crick Institute}
    • Durham - David Weinkove {Durham University} (via M. Barkoulas)
    • Leeds - Ian A. Hope {University of Leeds}
    • Bristol - Patricia Kuwabara {University of Bristol}
    • Cambridge - Olivia Casanueva {Babraham Institute}
    • Cambridge - Bill Schaeffer {MRC Laboratory of Molecular Biology}
    • Edinburgh - Maria Doitsidou {University of Edinburgh}

  • (9) Canada
    • Toronto, ON - Mei Zhen {Lunenfeld-Tanenbaum Research Institute}
    • Toronto, ON - Brent Derry {The Hospital for Sick Children}
    • Burnaby, BC - Michel Leroux {Simon Fraser University}
    • Trois-Rivières, QC - Patrick Narbonne {Université du Québec à Trois-Rivières}
    • Kingston, ON - Ian Chin-Sang {Queen s University}
    • Montreal, QC - Christian Rocheleau {Research Institute of the McGill University Health Centre}
    • Montreal, QC - Michael Hendricks via Christian Rocheleau {​Department of Biology, McGill University }
    • Vancouver, BC - Mark Edgley {University of British Columbia}
    • Burnaby, BC - Harald Hutter {Simon Fraser University}

  • (75) United States of America
    • Berkeley, CA - Michael Shapira {UC Berkeley, Department of Integrative Biology}
    • Davis, CA - Dan Starr {UC Davis}
    • Hayward, CA - Maria Gallegos {Cal State East Bay}
    • La Jolla, CA - Yishi Jin {UC San Diego}
    • La Jolla, CA - Emily Troemel {UC San Diego}
    • Los Angeles, CA - Sean Curran {University of Southern California}
    • Pasadena, CA - Paul Sternberg {California Institute of Technology}
    • Santa Clara, CA - Leilani Miller {Santa Clara University}
    • San Diego, CA - Arshad Desai {Ludwig Institute for Cancer Research, UCSD}
    • San Diego, CA - Supriya Srinivasan {The Scripps Research Institute}
    • San Francisco, CA - Noelle L Etoile {UCSF}
    • San Francisco, CA - Saul Kato {Sandler Neurosciences Center, UCSF}
    • Stanford, CA - Miriam Goodman {Stanford University}
    • Riverside, CA - Morris Maduro {University of California, Riverside}
    • Boulder, CO - Min Han {University of Colorado}
    • Hamden, CT - Alex De Lencastre {Quinnipiac University}
    • New Haven, CT - Shaul Yogev {Yale School of Medicine}
    • Willimantic, CT - Amy Groth {Eastern Connecticut State University}, via Erika Sorensen
    • Sarasota, FL - Katherine Walstrom {New College of FL}
    • North Chicago, IL - Hongkyun Kim {Rosalind Franklin University}
    • Chicago, IL - Dan Shaye {University of Illinois}
    • Chicago, IL - Chiou-Fen Chuang {University of Illinois at Chicago}
    • Crawfordsville, IN - Erika Sorensen {Wabash College}
    • Lawrence, KS - Brian D. Ackley {University of Kansas}
    • Manhattan, KS - Leah Radeke {Kansas State University}
    • Cambridge, MA - H. Robert (Bob) Horvitz {Massachusetts Institute of Technology}
    • Cambridge, MA - Steve Flavell {Massachusetts Institute of Technology}
    • Cambridge, MA - Susan Mango {MCB Harvard University}
    • Boston, MA - Alexander Soukas {MGH Harvard University}
    • Boston, MA - William Mair {Harvard T. H. Chan School of Public Health}
    • Boston, MA - Erin Cram {Northeastern University}
    • Waltham, MA - Piali Sengupta {Brandeis University} (via E. Yemini, Hobert lab)
    • Worcester, MA - Alexandra Byrne {University of Massachusetts Medical School}
    • Worcester, MA - Victor Ambros {University of Massachusetts Medical School}
    • Bethesda, MD - Kevin F O' Connell {NIH/National Institute of Diabetes & Digestive and Kidney Diseases}
    • Baltimore, MD - John Kim {Johns Hopkins University, Krieger School of Arts & Sciences}
    • Baltimore, MD - Yumi Kim {Johns Hopkins University, Krieger School of Arts & Sciences}
    • Ypsilanti, MI - Hannah Seidel {Eastern Michigan University}, via Erika Sorensen
    • Ann Arbor, MI - Shawn Xu {University of Michigan}
    • Ann Arbor, MI - Asim Beg {University of Michigan, Medical School}
    • Minneapolis, MN - Ann Rougvie {University of Minnesota}
    • Northfield, MN - Jennifer Wolff {Carleton College}
    • St Louis, MO - Stephen C. Pak {Washington University School of Medicine}
    • St Louis, MO - Zachary Pincus {Washington University School of Medicine}
    • Newark, NJ - Gal Haspel {New Jersey Institute of Technology}
    • Piscataway, NJ - Barth Grant {Rutgers University}
    • Piscataway, NJ - Martha Soto {Rutgers University}
    • Piscataway, NJ - Andrew Singson {‎Rutgers University, Waksman Institute of Microbiology }
    • Princeton, NJ - Andrew Leifer {Department of Physics, Princeton University}
    • Binghamton, NY - Howard C. Chang {Binghamton University}
    • New York, NY - Shai Shaham {Rockfeller University}
    • New York, NY - Oliver Hobert {Columbia University} (via S. Shaham)
    • New York, NY - Jeremy Nance {NYU School of Medicine}
    • New York, NY - Chris Li {CUNY-City College}
    • New York, NY - Itzik Mano (CUNY-City College)
    • New York, NY - Martin Chalfie {Columbia University}
    • New York, NY - Iva Greenwald {Columbia University}
    • New York, NY - Sevinc Ercan {NYU School of Medicine}
    • New York, NY - Gunsalus/Piano {NYU Center for Genomics and Systems Biology}
    • Bronx, NY - Hannes Buelow {Albert Einstein College of Medicine}
    • Ithaca, NY - Gunther Hollopeter {Cornell University}
    • Syracuse, NY - Sarah Hall {Syracuse University}
    • Syracuse, NY - Eleanor M. Maine {Syracuse University}
    • Gettysburg, PA - Jenifer Powell {Gettysburg College}
    • Villanova, PA - Matthew Youngman {Villanova University}
    • Philadelphia, PA - Gongyi Ren {Veterinary college of medicine, U Penn}
    • Oklahoma City, OK - Kenneth Miller {Oklahoma Medical Research Foundation}
    • Eugene, OR - Stephen Banse {University Of Oregon}
    • Portland, OR - Greg Hermann {Lewis and Clark College}
    • Providence, RI - Saba Baskoylu {Brown University}
    • Nashville, TN - David M. Miller {Vanderbilt University}
    • Harrogate, TN - Melissa Henderson {Lincoln Memorial University}
    • Houston, TX - Natasha Kinenko {Rice University}
    • San Antonio, TX - Alfred Fisher {University of Texas Health Science Center at San Antonio}
    • Salt Lake City, UT - Ofer Rog {University of Utah}
    • Milwaukee, WI - Chris Quinn {University of Wisconsin}
    • Madison, WI - Judith Kimble {University of Wisconsin}, via Erika Sorensen
    • Madison, WI - Ahna R. Skop {University of Wisconsin}

  • (1) Israel
    • Tel Aviv - Oded Rechavi {Tel-Aviv University}

  • (4) Australia
    • Clayton - Brent Neumann {Monash University}
    • Clayton - Roger Pocock {Monash University} (via B. Neurmann)
    • Clayton - Jie Liu {Monash University} (via B. Neurmann)
    • Brisbane - Massimo A. Hilliard {University of Queensland} (via B. Neumann)

  • (3) China
    • Wuhan - Anbing Shi {Huazhong University of Science and Technology}
    • Shanghai - Zhiyong Shao {Institute of Brain Science, Fudan University}
    • Shanghai - Yan Zou {ShanghaiTech University}

  • (2) Taiwan
    • Tainan - Chang-Shi Chen {National Cheng Kung University}
    • Taipei - Shih-Peng Chan {National Taiwan University} via Chang-Shi Chen

  • (1) Hong Kong
    • Hong Kong - Ho Yi Mak {Hong Kong University of Science and Technology}

  • (7) Japan
    • Fukuoka - Takeshi Ishihara {Kyushu university}
    • Yokohama - Ken-ichi Ogura {Yokohama City University School of Medicine}
    • Kobe - Shuichi Onami {RIKEN} via Takeshi Ishihara
    • Kyoto - Peter Carlton {Kyoto University} via Hitoshi Sawa
    • Mishima - Hitoshi Sawa {National Institute of Genetics} via Ken-ichi Ogura
    • Osaka - Yashushi Okamura {Osaka University}
    • Tsukuba, Ibaraki - Motomichi Doi {AIST}

  • (2) South Korea
    • Seoul - Junho Lee {Seoul National University}
    • Cheongam-ro - Seung-Jae Lee {POSTECH} via Junho Lee

  • (1) India
    • Mohali - Yogesh Dahiya, Kavita Babu lab {IISER}

Plasmids from our paper.

pSEM87 : wrmScarlet cDNA sequence. Download sequence here.
pSEM89 : SL2::wrmScarlet artificial operon. Download sequence here.
pSEM90 : wrmScarlet-I cDNA sequence. Download sequence here.
pMD8 : sgRNA targeting the d10 site. Download sequence here.
pPT2 : sgRNA expression vector. Download sequence here.

"Right-click" on the file name and use "Save to Link As..." to save the file on your hard drive. This annotated sequence file is in Genbank format and can be opened with most software packages. We like ApE and benchling.com.

wrmScarletComparison
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