Live Imaging of mRNA Synthesis in Drosophila.

نویسندگان

  • Hernan G Garcia
  • Thomas Gregor
چکیده

mRNA synthesis is one of the earliest readouts of the activity of a transcribed gene, which is of particular interest in the context of metazoan cell fate specification. These processes are intrinsically dynamic and stochastic, which makes in vivo single-cell measurements inevitable. Here, we present the application of a technology that has been widely used in single celled organisms to measure transcriptional activity in developing embryos of the fruit fly Drosophila melanogaster. The method allows for quantification of instantaneous polymerase occupancy of active gene loci and thereby enables the development and testing of models of gene regulation in development.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Preparing Individual Drosophila Egg Chambers for Live Imaging

Live cell imaging is an important technique applied to a number of Drosophila tissues used as models to investigate topics such as axis specification, cell differentiation and organogenesis (1). Correct preparation of the experimental samples is a crucial, often neglected, step. The goal of preparation is to ensure physiological relevance and to establish optimal imaging conditions. To maintain...

متن کامل

Retraction Notice to: Assembly of Endogenous oskar mRNA Particles for Motor-Dependent Transport in the Drosophila Oocyte

(Cell 139, 983–998; November 25, 2009) In this paper, we used cryoimmuno-electron microscopy and live-cell imaging to investigate the sequential assembly of oskarmRNA into an mRNP competent for transport from the Drosophila nurse cells to the oocyte posterior pole. We have recently identified instances in all of the figures where the cryoimmuno-EM data were inappropriately manipulated by the fi...

متن کامل

A late phase of germ plasm accumulation during Drosophila oogenesis requires lost and rumpelstiltskin.

Asymmetric mRNA localization is an effective mechanism for establishing cellular and developmental polarity. Posterior localization of oskar in the Drosophila oocyte targets the synthesis of Oskar to the posterior, where Oskar initiates the assembly of the germ plasm. In addition to harboring germline determinants, the germ plasm is required for localization and translation of the abdominal det...

متن کامل

Imaging Single-mRNA Localization and Translation in Live Neurons

Local protein synthesis mediates precise spatio-temporal regulation of gene expression for neuronal functions such as long-term plasticity, axon guidance and regeneration. To reveal the underlying mechanisms of local translation, it is crucial to understand mRNA transport, localization and translation in live neurons. Among various techniques for mRNA analysis, fluorescence microscopy has been ...

متن کامل

bicoid mRNA localises to the Drosophila oocyte anterior by random Dynein-mediated transport and anchoring

bicoid mRNA localises to the Drosophila oocyte anterior from stage 9 of oogenesis onwards to provide a local source for Bicoid protein for embryonic patterning. Live imaging at stage 9 reveals that bicoid mRNA particles undergo rapid Dynein-dependent movements near the oocyte anterior, but with no directional bias. Furthermore, bicoid mRNA localises normally in shot2A2, which abolishes the pola...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Methods in molecular biology

دوره 1649  شماره 

صفحات  -

تاریخ انتشار 2018