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. 2024 Mar 12:2024:10.17912/micropub.biology.001150.
doi: 10.17912/micropub.biology.001150. eCollection 2024.

Characterizing a standardized BioPart for BAG-specific expression in C. elegans

Xinglin Qian  1 , Sarah AlHarbi  1 , Christian Frøkjær-Jensen  1
Affiliations

Characterizing a standardized BioPart for BAG-specific expression in C. elegans

Xinglin Qian et al. MicroPubl Biol. .

Abstract

Biological parts (BioParts) are modular and standardized DNA sequences that encode biological functions and contribute to the efficient biological engineering of complex systems. Here, we characterize a short BioPart (P flp-17 , 300 bp) for bright multicopy and single-copy BAG-specific expression starting from the gastrula stage in hermaphrodite and male C. elegans . We have generated standardized P flp-17 cloning vectors for BAG-specific gfp and mScarlet expression compatible with extra-chromosomal arrays and for single-copy transgene insertion. The short P flp-17 promoter is easy to generate by gene synthesis and has been incorporated into our online transgene design tool (www.wormbuilder.org/transgenebuilder ) .

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Conflict of interest statement

The authors declare that there are no conflicts of interest present.

Figures

Figure 1. <b> Expression and Characterization of P <i>flp-17 </i> in BAG Neurons of <i>C. elegans</i> </b>
Figure 1. Expression and Characterization of P flp-17 in BAG Neurons of C. elegans
A . In hermaphrodites (top panel) and males (bottom panel) a P flp-17 :: mScarlet transgene is expressed exclusively in BAG neurons (20x magnification, scale bars = 100 µm). B . Single-copy P flp-17 :: mScarlet expression in a hermaphrodite (20x magnification, scale bar = 20 µm). C . flp-17 promoter characteristics. CenGEN data from https://cengen.shinyapps.io/CengenApp/ and https://cengen.textpressolab.com/. D . Schematic of standard mScarlet and gfp vectors for BAG-specific expression and identification. Bsa I restriction sites allow exchanging the fluorescent reporter for a gene of interest by Golden Gate Assembly; Apa I (N-terminal) and Sma I (C-terminal) allow fluorophore fusions. The vectors are compatible with single-copy insertions (MosTI) and expression from extra-chromosomal arrays. E . Illustration of the transgenebuilder workflow. The short 300 bp P flp-17 was added as a standard promoter in our online tool for optimizing transgene design and gene synthesis (www.wormbuilder.org/transgenebuilder).
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Grants and funding

This work was funded by a competitive research grant from the KAUST Office of Research Administration (CRG9 URF/1/4388-01-01).

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