Hello, I'm Abby, I recently completed my 3rd year studying Biomedical Sciences at the University of Warwick. This blog documents my Undergraduate Research Support Scheme (URSS), where for 10 weeks between July and September I explored a developmental biology lab working with Caenorhabditis elegans (C. elegans) and their embryos. This project was taken place under Dr Erik Griffin, and I have been working alongside my supervisors Dr Bernardo Chapa y Lazo and Dr Sally Adams.

With only one prior wet lab experience, my 3rd year undergraduate research project, I aim to deepen my understanding of the realities and responsibilities of working in a research lab.

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About my project:

The centrosome is​ a crucial cellular structure involved in mitosis, the division of cells (1). These structures are able to produce mitotic spindles which allow the equal separation of chromosomes to each daughter cell. Beyond spindle assembly, centrosomes, an essential structure during cellular division, plays a key role in cell polarity and intracellular organisation (2). Within centrosomes are miniture structures, centrioloes. Like the centrosome, these structures form and organise microtubules providing a structural foundation for spindle formation during cell division (3).

Despite there being a range of literature on centrosomal function, little is known about exact mechanism for centrosome formation. In particular, my project focuses on three proteins in the centrosome. These are SAS-4, SAS-6 and SPD-2, chosen by my principle investigator, Dr Erik Griffin.

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SAS-4, a key scaffolding protein in the centrioles, recruits other strucutral proteins and stabilises the extracellular matrix (4). SAS-6, another core centriolar protein, responsible for centriolar symmetry and structural integrity of the centrioles (5). The final protein, SPD-2 is important for the recruitment of pericentriolar material (PCM) (6).

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A key question driving this work is whether these centriolar proteins are synthesised locally at the centrosome itself, or are they synthesised elsewhere and are transported to the site of centrosomal assembly. Not only is this process key to life, this process occurs rapidly, within a few minutes of our model organism C. elegans. With a divison time of 20 minutes in the early embryo, how are the centrosomes coordinated and form before the 20 minute time limit.

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These worms provide key biological insights into mammalian cell divison and development, with a rapid division rate, self-fertilising and easy genetic manipulation (7). My project involves the synthesis of and cloning of GFP tags, allowing the fluoresence and visualisation of the proteins of interest and inserting them into the worm genotype. This project involves the use of sun-tags to allow the live imaging of mRNA synthesis of the genes coding for SAS-4, SAS-6 and SPD-2.

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First, I plan to clone homology arms into the GFP tagged template using golden gate cloning. These clones will then be purified and sent for sequencing to confirm the correct homology arms were cloned into the tags. These clones will then be amplified and purified for injection into our C. elegans strains. The F1 progeny of the injected worms will be picked, grown and screened for the template. The F1 progeny heterozygous for the insert would then chosen and F2 progeny from this worm will again be picked, grown and screened for the homozygous insert. Once a positive strain is identified, this strain's embryos will be studied and visualised under a microscope to see the live transcription of our centriolar proteins, allowing us to identify where these proteins are made.