Literature DB >> 24852212

Single and dual drug selection for transgenes following bombardment of Caenorhabditis species.

Jennifer I Semple1, Ben Lehner2.   

Abstract

The use of drugs and drug resistance genes is a powerful method to select for the presence of a transgene. Unlike methods that require the complementation of a genetic mutation, this system can be used on any genetic background. Drug selection does not require extensive manipulation or costly equipment, yet it is very rapid and can achieve extremely high efficiency, selecting a small number of transgenic worms from among millions of non-transgenic worms. Introducing integrated transgenes into Caenorhabditis elegans by microparticle bombardment represents just such a challenge. Here we describe in detail the protocol we have developed for dual-drug selection in liquid with puromycin and G418 which works well in a variety of Caenorhabditis species. We also show that single drug selection with only puromycin or only G418 is effective in C. elegans. The growing number of drug selection markers that have been adapted to C. elegans are an important addition to the genetic toolkit at our disposal.
Copyright © 2014 Elsevier Inc. All rights reserved.

Entities:  

Keywords:  Antibiotic; C. brenneri; C. briggsae; C. elegans; C. remanei; Drug selection; G418; Microparticle bombardment; Puromycin; Transgenesis

Mesh:

Substances:

Year:  2014        PMID: 24852212     DOI: 10.1016/j.ymeth.2014.05.004

Source DB:  PubMed          Journal:  Methods        ISSN: 1046-2023            Impact factor:   3.608


  5 in total

1.  Expanding the C. elegans toolbox into a toolshed.

Authors:  Arjumand Ghazi; Judith Yanowitz; Gary A Silverman
Journal:  Methods       Date:  2014-08-01       Impact factor: 3.608

2.  Transgenesis by microparticle bombardment for live imaging of fluorescent proteins in Pristionchus pacificus germline and early embryos.

Authors:  Satoshi Namai; Asako Sugimoto
Journal:  Dev Genes Evol       Date:  2018-01-20       Impact factor: 0.900

3.  Transgenesis of the gonochoristic nematode Caenorhabditis inopinata by microparticle bombardment with hygromycin B selection.

Authors:  Shun Oomura; Kenji Tsuyama; Nami Haruta; Asako Sugimoto
Journal:  MicroPubl Biol       Date:  2022-05-05

4.  Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping.

Authors:  Cody L Gilleland; Adam T Falls; James Noraky; Maxwell G Heiman; Mehmet F Yanik
Journal:  Genetics       Date:  2015-07-10       Impact factor: 4.562

5.  Homeodomain-Interacting Protein Kinase (HPK-1) regulates stress responses and ageing in C. elegans.

Authors:  Slavica Berber; Mallory Wood; Estelle Llamosas; Priya Thaivalappil; Karen Lee; Bing Mana Liao; Yee Lian Chew; Aaron Rhodes; Duygu Yucel; Merlin Crossley; Hannah R Nicholas
Journal:  Sci Rep       Date:  2016-01-21       Impact factor: 4.379

  5 in total

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