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This cruise was the second of in total four seasonal cruises with RV Kronprins Haakon in 2019/20 focusing on biology in the project Arven etter Nansen (AeN). This seasonal cruise was named Q4 (Q4= 4th quarter of the year) investigating in total 17 stations of the established AeN transect along 34 E in the Northern Barents Sea and adjacent Arctic Basin from 76 to 82°N (see Fig. 1 below). The cruise addressed objectives of the research foci in RF1 on Physical drivers, RF2 on Human drivers, RF3 on the living Barents Sea and RA-C Technology and method development, and collected a multitude of data along the Nansen Legacy transect which was ice covered except the southernmost station P1. In addition to in situ sampling, on board experiments were conducted to quantify biological processes, rates and interactions that will also be important feeds into modeling work and projections in RF4 The future Barents Sea. The cruise took a variety of continuous ship measurements (Weather station, EK80, EM203, ADCP, thermosalinograph, pCO2 underway) as well as station measurements such as CTD with water samples, biological sampling of the benthos (box corer, benthic trawl), water column (multinet, MIK net, macrozooplankton trawl and many other smaller nets) and sea ice (snow, ice cores, water just underneath sea ice). In addition, experimental work (respiration, grazing and egg production) was conducted in the ship’s laboratories. The chemistry team onboard measured oxygen, nutrients and pH from standard depths on most CTD stations and sea ice samples. The cruise started in Longyearbyen and ended in Tromsø (28.11.-17.12.2019). The sampling began at the deep (>3000 m) northernmost station of the transect, Stn. P7, and continued along the southward transect until station P1, in open water and Atlantic dominated water masses. During the expedition the Barents Sea was characterized by a relatively large sea ice cover with consolidated sea ice all the way from P7 to P2. The Polar Front was located just north of P1. All process stations were sampled (P7-P1) as well as two ice stations: one close to P7 ad one close to P5. At the southernmost station P1, stormy weather challenged sampling, but most tasks were in the end accomplished except of deploying the box corer, sediment trap and the AUV. These operations were considered too challenging due to strong drift and ship movement, and it was not safe to conduct small boat operations. Challenges with the box corer was also experienced at the deep station P7 due to technical issues. In the end, most work was accomplished despite challenging weather, sea ice conditions and some technical issues making this cruise successful in gaining new important knowledge about the Northern Barents Sea in the polar night season which is extremely poorly studied. The overall high biological activity and biomass at this time of the year, November-December, was surprising for most of us.

Summary The CRISPR/Cas9 system is an RNA‐guided sequence‐specific genome editing tool, which has been adopted for single or multiple gene editing in a wide range of organisms. When working with gene families with functional redundancy, knocking out multiple genes within the same family may be required to generate a phenotype. In this study, we tested the possibility of exploiting the known tolerance of Cas9 for mismatches between the single‐guide RNA (sgRNA) and target site to simultaneously introduce indels in multiple homologous genes in the marine diatom Phaeodactylum tricornutum. As a proof of concept, we designed two sgRNAs that could potentially target the same six light‐harvesting complex (LHC) genes belonging to the LHCF subgroup. Mutations in up to five genes were achieved simultaneously using a previously established CRISPR/Cas9 system for P. tricornutum. A visible colour change was observed in knockout mutants with multiple LHCF lesions. A combination of pigment, LHCF protein and growth analyses was used to further investigate the phenotypic differences between the multiple LHCF mutants and WT. Furthermore, we used the two same sgRNAs in combination with a variant of the existing Cas9 where four amino acids substitutions had been introduced that previously have been shown to increase Cas9 specificity. A significant reduction of off‐target editing events was observed, indicating that the altered Cas9 functioned as a high‐fidelity (HiFi) Cas9 nuclease.