Advanced non-viral gene delivery experiments often require co-delivery of multiple nucleic acids. Therefore, the availability of reliable and robust co-transfection methods and defined selection criteria for their use in, e.g., expression of multimeric proteins or mixed RNA/DNA delivery is of utmost importance. Here, we investigated different co- and successive transfection approaches, with particular focus on in vitro transcribed messenger RNA (IVT-mRNA). Expression levels and patterns of two fluorescent protein reporters were determined, using different IVT-mRNA doses, carriers, and cell types. Quantitative parameters determining the efficiency of co-delivery were analyzed for IVT-mRNAs premixed before nanocarrier formation (integrated co-transfection) and when simultaneously transfecting cells with separately formed nanocarriers (parallel co-transfection), which resulted in a much higher level of expression heterogeneity for the two reporters. Successive delivery of mRNA revealed a lower transfection efficiency in the second transfection round. All these differences proved to be more pronounced for low mRNA doses. Concurrent delivery of siRNA with mRNA also indicated the highest co-transfection efficiency for integrated method. However, the maximum efficacy was shown for successive delivery, due to the kinetically different peak output for the two discretely operating entities. Our findings provide guidance for selection of the co-delivery method best suited to accommodate experimental requirements, highlighting in particular the nucleic acid dose-response dependence on co-delivery on the single-cell level.A novel white-colored, aerobic, Gram-stain-positive, rod-shaped bacterium, designated strain DB0629T was isolated from a motor car evaporator core collected in South Korea. Strain DB0629T grew at 10-35 °C, pH 6.0-9.0, and 0-5.0% (w/v) NaCl concentration. The 16S rRNA gene sequence analysis showed that strain DB0629T belonged to the genus Nakamurella, with the nearest phylogenetic neighbor being Nakamurella lactea DSM 19367T (97.6% sequence similarity). The strain comprised diphosphatidylglycerol and phosphatidylinositol as the main polar lipids; MK-8(H4) as a sole respiratory quinone; meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan and anteiso-C150, anteiso-C170, iso-C160, iso-C150, and C160 as the major fatty acids. The average nucleotide identity (ANI) and in silico DNA-DNA hybridization values between strain DB0629T and N. lactea DSM 19367T were 74.9% and 20.8%, respectively, which were below the threshold values of 95% and 70%, respectively. The DNA G + C content was 69.5 mol%. Based on the polyphasic taxonomic data, the novel species Nakamurella aerolata sp. nov. is proposed with the type strain DB0629T (= KCTC 72726T = NBRC 114624T).A light yellow-colored, Gram-stain-negative, aerobic, oxidase- and catalase-positive, flagellated bacterium with motility, designated as strain AE3T was isolated from soil. Cells of strain AE3T are rod-shaped, and the colonies are round and convex. Phylogenetic analysis based on 16S rRNA gene sequence revealed that strain AE3T forms a lineage within the genus Sphingomonas of the family Sphingomonadaceae and is most closely related to Sphingomonas edaphi KCTC 62107 T (98.6%), Sphingomonas oryziterrae KCTC 22476 T (97.9%), and Sphingomonas jaspsi DSM 18422 T (97.4%). The growth of the strain AE3T was observed under 18-42 °C (optimum, 37 °C), pH 6.0-9.0 (optimum, pH 6.5-7.0), and in the absence of NaCl. Strain AE3T contains Q-10 as a predominant respiratory quinone, and the major fatty acids are C171 ω6c, summed feature 8 (C181 ω7c), and summed feature 3 (C161 ω7c and/or C161 ω6c). The major polar lipids are sphingoglycolipids, unidentified phospholipids, and phosphatidylethanolamine. The DNA G + C content of strain AE3T is 63.6 mol%. The nearly complete genome of strain AE3T consists of 2.2 Mbp, (2,168 total protein-coding genes, 45 tRNAs, 4 ncRNAs, and 3 rRNAs). Genomic taxonomy analysis demonstrates that the novel strain has  less then  75.9% average nucleotide identity value, and also shows  less then  24.9% in silico DNA-DNA hybridization value compared to related taxa, which clearly separates strain AE3T from other species of the genus Sphingomonas with values below the thresholds for species delineation. Based on phenotypic, genotypic, and phylogenetic analyses, strain AE3T represents a novel species of the genus Sphingomonas, for which the name Sphingomonas xanthus sp. nov. is proposed. The type strain of Sphingomonas xanthus is AE3T (= KCTC 620106 T = JCM 32376 T). Part 1 of this two-part, open-label, Phase 1 study (NCT03233139) assessed the safety, tolerability, pharmacokinetics, immunogenicity, andclinical activity of cemiplimab in Japanese patients with advanced malignancies. Patients received cemiplimab 250mg (n = 6) or 350mg (n = 7) every 3weeks intravenously for up to 108weeks in Part 1. Tumor responses were assessed by investigators every 9weeks using the Response Evaluation Criteria in Solid Tumors version 1.1. Of 13 patients enrolled, median age was 62years (range 33-75) and eight patients were female. Median duration of cemiplimab exposure was 13.1weeks (range 3.0‒113.6). At the time of data cut-off, 11 patients (84.6%) had discontinued treatment (majority due to disease progression n = 8, 61.5%). trans-4-Hydroxytamoxifen The most common treatment-emergent adverse events (TEAEs) of any grade were contact dermatitis, rash, and viral upper respiratory tract infection (each n = 3, 23.1%). Five grade ≥ 3 TEAEs were reported in four patients autoimmune colitis, dehydration, hyponatremia, hypophosphatemia, and muscular weakness. No dose-limiting toxicities were reported and no TEAEs led to death. Cemiplimab concentrations in serum were consistent with previously reported pharmacokinetic characteristics of cemiplimab. No anti-drug antibodies were detected in serum. Objective response rate [ORR; complete response + partial response (PR)] was 30.8% (four PR) and disease control rate [ORR + stable disease (SD)] was 46.2% (6/13; two SD). Cemiplimab exhibited antitumor activity in Japanese patients with advanced malignancies. The safety profile was comparable to those previously reported for cemiplimab and other PD-1 inhibitors. NCT03233139 at ClinicalTrials.gov.NCT03233139 at ClinicalTrials.gov.