Genetic similarity of biological samples to counter bio-hacking of DNA-sequencing functionality.

We present the work towards strengthening the security of DNA-sequencing functionality of future bioinformatics systems against bio-computing attacks. Recent research has shown how using common tools, a perpetrator can synthesize biological material, which upon DNA-analysis opens a cyber-backdoor for the perpetrator to hijack control of a computational resource from the DNA-sequencing pipeline. As DNA analysis finds its way into practical everyday applications, the threat of bio-hacking increases. Our wetlab experiments establish that malicious DNA can be synthesized and inserted into E. coli, a common contaminant. Based on that, we propose a new attack, where a hacker to reach the target hides the DNA with malicious code on common surfaces (e.g., lab coat, bench, rubber glove). We demonstrated that the threat of bio-hacking can be mitigated using dedicated input control techniques similar to those used to counter conventional injection attacks. This article proposes to use genetic similarity of biological samples to identify material that has been generated for bio-hacking. We considered freely available genetic data from 506 mammary, lymphocyte and erythrocyte samples that have a bio-hacking code inserted. During the evaluation we were able to detect up to 95% of malicious DNAs confirming suitability of our method.

The use of clade-specific PCR assays to identify novel nitrilase genes from environmental isolates.

Nitrilase enzymes (EC 3.5.5.1) are responsible for the direct hydration of nitriles to their corresponding carboxylic acids and ammonia. The utilization of nitrilase enzymes in biocatalysis toward bio-pharmaceuticals and industrial applications facilitates the move towards green chemistry. The body of research presented describes a novel clade-specific touchdown PCR protocol for the detection of novel nitrilase genes. The presented study identified partial sequences of 15 novel nitrilase genes across 7 genera, with partial DNA sequence homology (%) displayed across an additional 16 genera. This research will prove valuable in the screening of microorganisms for the identification of novel clade-specific nitrilase genes, with predicted enantioselective profiles as determined by their clade characterizations.

Real-time PCR detection of aldoxime dehydratase genes in nitrile-degrading microorganisms.

Aldoxime dehydratase catalyses the conversion of aldoximes to their corresponding nitriles. Utilization of the aldoxime-nitrile metabolising enzyme pathway can facilitate the move towards a greener chemistry. In this work, a real-time PCR assay was developed for the detection of aldoxime dehydratase genes in aldoxime/nitrile metabolising microorganisms which have been purified from environmental sources. A conventional PCR assay was also designed allowing gene confirmation via sequencing. Aldoxime dehydratase genes were identified in 30 microorganisms across 11 genera including some not previously shown to harbour the gene. The assay displayed a limit of detection of 1 pg/µL DNA or 7 CFU/reaction. This real-time PCR assay should prove valuable in the high-throughput screening of micro-organisms for novel aldoxime dehydratase genes towards pharmaceutical and industrial applications.