De novo assembly and annotation of the CHOZN® GS-/- genome supports high-throughput genome-scale screening.

Chinese hamster ovary (CHO) cells have been used as the industry standard for the production of therapeutic monoclonal antibodies for several decades. Despite significant improvements in commercial-scale production processes and media, the CHO cell has remained largely unchanged. Due to the cost and complexity of whole-genome sequencing and gene-editing it has been difficult to obtain the tools necessary to improve the CHO cell line. With the advent of next-generation sequencing and the discovery of the CRISPR/Cas9 system it has become more cost effective to sequence and manipulate the CHO genome. Here, we provide a comprehensive de novo assembly and annotation of the CHO-K1 based CHOZN® GS-/- genome. Using this platform, we designed, built, and confirmed the functionality of a whole genome CRISPR guide RNA library that will allow the bioprocessing community to design a more robust CHO cell line leading to the production of life saving medications in a more cost-effective manner.

Chemical immobilisation of dhole (Cuon alpinus), Indian jackal (Canis aureus indicus) and Indian wolf (Canis lupus pallipes) with ketamine hydrochloride-xylazine hydrochloride.

Maintaining wild animals in captivity often requires chemical immobilisation to achieve various diagnostic, surgical and management interventions. Four dholes, two Indian grey wolves and four Indian jackals were immobilised using ketamine-xylazine combination for either medical or management interventions. Based on the estimated body weight, canids were darted upon with 6-8 mg kg-1 ketamine and 0.7-1.14 mg kg-1 xylazine. Initial signs of drug effect included decreased mentation and progressive ataxia followed by recumbency. The mean ± SD of induction time was 14.25 ± 2.75 (range: 11-17 min), 11 ± 3.16 (range: 8-15 min) and 15.5 ± 3.54 (range: 13-18 min) in dhole, Indian jackal and Indian wolf, respectively. Hyperthermia was initially observed in all the jackals and dholes, whereas rectal temperature in wolves remained well within the normal range for canids. The mean duration of anaesthesia was 31 ± 8.83 (range: 23-43 min), 32.5 ± 5.32 (range: 26-39 min) and 30.5 ± 7.78 (range: 25-36 min) in dhole, Indian jackal and Indian wolf, respectively, with subsequent smooth and uneventful recovery in all the cases. The observations made during immobilisation procedures in this work suggest that chemical immobilisation of captive dhole, Indian wolf and Indian jackal with 6-8 mg kg-1 ketamine and 1 mg kg-1 xylazine is effective and safe for routine management and medical interventions in these species provided body temperature is closely monitored and corrected as appropriate.