An optimized QF-binary expression system for use in zebrafish.

Developmental Biology
Jason BurgessBret J Pearson

Abstract

The zebrafish model organism has been of exceptional utility for the study of vertebrate development and disease through the application of tissue-specific labelling and overexpression of genes carrying patient-derived mutations. However, there remains a need for a binary expression system that is both non-toxic and not silenced over animal generations by DNA methylation. The Q binary expression system derived from the fungus Neurospora crassa is ideal, because the consensus binding site for the QF transcription factor lacks CpG dinucleotides, precluding silencing by CpG-meditated methylation. To optimize this system for zebrafish, we systematically tested several variants of the QF transcription factor: QF full length; QF2, which lacks the middle domain; QF2w, which is an attenuated version of QF2; and chimeric QFGal4. We found that full length QF and QF2 were strongly toxic to zebrafish embryos, QF2w was mildly toxic, and QFGal4 was well tolerated, when injected as RNA or expressed ubiquitously from stable transgenes. In addition, QFGal4 robustly activated a Tg(QUAS:GFPNLS) reporter transgene. To increase the utility of this system, we also modified the QF effector sequence termed QUAS, which consists of five copies of the QF...Continue Reading

References

Dec 5, 1995·Proceedings of the National Academy of Sciences of the United States of America·M L HedleyT Maniatis
Mar 12, 1999·Mechanisms of Development·N Scheer, J A Campos-Ortega
May 11, 2000·Proceedings of the National Academy of Sciences of the United States of America·D Szüts, M Bienz
Dec 6, 2005·Mechanisms of Development·Vincent T Cunliffe, Patrizia Casaccia-Bonnefil
Apr 4, 2006·Nature Neuroscience·Sen-Lin Lai, Tzumin Lee
Mar 21, 2007·Nature Methods·Ethan K ScottHerwig Baier
Oct 17, 2007·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Kristen M KwanChi-Bin Chien
Jan 19, 2008·Proceedings of the National Academy of Sciences of the United States of America·Kazuhide AsakawaKoichi Kawakami
Jun 12, 2008·Developmental Biology·Alexander Emelyanov, Serguei Parinov
May 13, 2009·Genetics·Mary G GollMarnie E Halpern
Jul 25, 2009·Proceedings of the National Academy of Sciences of the United States of America·Martin DistelReinhard W Köster
Sep 2, 2010·Proceedings of the National Academy of Sciences of the United States of America·Ryohei YagiKonrad Basler
Jan 13, 2011·Developmental Biology·Courtney M AkitakeMary G Goll
Jun 30, 2012·Nature Methods·Johannes SchindelinAlbert Cardona
Jun 25, 2013·Methods : a Companion to Methods in Enzymology·Abhignya SubediMarnie E Halpern
Oct 11, 2014·Nucleic Acids Research·Eva K BrinkmanBas van Steensel
Jan 13, 2015·Nature Methods·Olena RiabininaChristopher J Potter
Mar 10, 2016·Genome Research·Vincenzo Di DonatoFilippo Del Bene
Mar 24, 2016·Developmental Cell·Kazuyuki HoshijimaDavid Jonah Grunwald
May 12, 2016·Advances in Experimental Medicine and Biology·Marie Mayrhofer, Marina Mione
Jul 23, 2016·Methods in Cell Biology·A Ghosh, M E Halpern
Dec 1, 2017·BMC Bioinformatics·Curtis T RuedenKevin W Eliceiri
May 21, 2019·Nucleic Acids Research·Kornel LabunEivind Valen

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.