Rosell, F. | bioRxiv Science

bioRxiv2026

Process-based model predicts seasonal variation in eDNA transport - a case study on Eurasian beavers in a small river

Robust methods to monitor species distributions are vital to ensuring successful conservation strategies, particularly in the case of conservation translocations. Environmental DNA (eDNA) from water samples is a cost-effective method to monitor species distributions without physical capture or disturbance. However, eDNA is vulnerable to long-distance transport depending on the hydrological and environmental characteristics which can lead to spatially false positives and ultimately inaccurate species distributions. Recently, the development of particle transport models has allowed researchers to integrate hydrological and environmental variables to predict how far eDNA will transport from a source point. Here, eDNA samples (n=218) were collected and quantified using digital PCR (dPCR) to study monthly changes in Eurasian beaver (Castor fiber) eDNA concentrations downstream of an enclosure which contained 4 - 5 beavers located in Scotland. The shortest eDNA transport distances (< 2 km) were observed in the summer which correlated with the lowest flows and highest temperatures. In contrast, throughout the winter eDNA was consistently detected up to 5.8 km downstream correlating with the highest discharge and lowest temperature. The eDNA transport model reliably reproduced the decrease in eDNA concentrations downstream of the enclosure, however there were challenges surrounding stream-specific decay rates following a confluence. To study localised species distributions, samples should be collected during summer low flow conditions. Conversely, to maximise species detections sampling should be conducted in winter which had the longest eDNA transport and highest detectability.

ecology↗

bioRxiv2018

Optimizing guide RNA selection and CRISPR/Cas9 methodology for efficient generation of deletions in C. elegans.

The Caenorhabditis elegans Gene Knockout (KO) Consortium is tasked with obtaining null mutations in each of the more than 20,000 open reading frames (ORFs) of this organism. To date, approximately15,000 ORFs have associated putative null alleles. A directed approach using CRISPR/Cas9 methodology is the most promising technique to complete the task. While there has been substantial success in using CRISPR/Cas9 in C. elegans, there has been little emphasis on optimizing the method for generating large insertions/deletions in this organism. To enhance the efficiency of using CRISPR/Cas9 to generate gene knockouts in C. elegans we have developed an online species-specific guide RNA selection tool (http://genome.sfu.ca/crispr). When coupled with previously developed selection vectors, optimization for homology arm length, and the use of purified Cas9 protein, we demonstrate a robust, efficient and effective protocol for generating deletions. Debate and speculation in the larger scientific community about off- target effects due to non-specific Cas9 cutting has prompted us to investigate through whole genome sequencing the occurrence of single nucleotide variants and indels accompanying targeted deletions. We did not detect any off-site variants above the natural spontaneous mutation rate and therefore conclude this modified protocol does not generate off-target events to any significant degree in C. elegans.

genetics↗