The data in this article are associated with the research paper “GigaAssay – an adaptable high-throughput saturation mutagenesis assay” [1]. The raw data are sequence reads of HIV-1 Tat cDNA amplified from cellular genomic DNA in a new single-pot saturation mutagenesis assay designated the “GigaAssay”. A bioinformatic pipeline and parameters used to analyze the data. Raw, processed, analyzed, and filtered data are reported. The data is processed to calculate the Tat-driven transcription activity for cells with each possible single amino acid substitution in Tat. This data can be reused to interpret Tat intermolecular interactions and HIV latency. This is one of the largest and most complete datasets regarding the impact of amino acid substitutions within a single protein on a molecular function.

Double-strand breaks (DSB) are one of the most lethal forms of DNA damage that, if left unrepaired, can lead to genomic instability, cellular transformation, and cell death. In this work, we examined how repair of transcription activator-like effector nuclease (TALEN)-induced DNA damage was altered when knocking out, or inhibiting a function of, two DNA repair proteins, XRCC4 and MRE11, respectively. We developed a fluorescent reporter assay that uses TALENs to introduce DSB and detected repair by the presence of GFP fluorescence. We observed repair of TALEN-induced breaks in the XRCC4 knockout cells treated with mirin (a pharmacological inhibitor of MRE11 exonuclease activity), albeit with ~40% reduced efficiency compared to normal cells. Editing in the absence of XRCC4 or MRE11 exonuclease was robust, with little difference between the indel profiles amongst any of the groups. Reviewing the transcriptional profiles of the mirin-treated XRCC4 knockout cells showed 307 uniquely differentially expressed genes, a number far greater than for either of the other cell lines (the HeLa XRCC4 knockout sample had 83 genes, and the mirin-treated HeLa cells had 30 genes uniquely differentially expressed). Pathways unique to the XRCC4 knockout+mirin group included differential expression of p53 downstream pathways, and metabolic pathways indicating cell adaptation for energy regulation and stress response. In conclusion, our study showed that TALEN-induced DSBs are repaired, even when a key DSB repair protein or protein function is not operational, without a change in indel profiles. However, transcriptional profiles indicate the induction of unique cellular responses dependent upon the DNA repair protein(s) hampered.

Motor vehicle crashes are one of the leading causes of death among teenagers. Many of these deaths are due to preventable causes, including impaired and distracted driving. You Drink, You Drive, You Lose (YDYDYL) is a prevention program to educate high school students about the consequences of impaired and distracted driving. YDYDYL was conducted at a public high school in Southern Nevada in March 2020. A secondary data analysis was conducted to compare knowledge and attitudes of previous participants with first-time participants. Independent-samples-t test and χ² test/Fisher’s exact test with post-contingency analysis were used to compare pre-event responses between students who had attended the program one year prior and students who had not. Significance was set at p < 0.05. A total of 349 students participated in the survey and were included for analysis; 177 had attended the program previously (50.7%) and 172 had not (49.3%). The mean age of previous participants and first-time participants was 16.2 (SD ± 1.06 years) and 14.9 (SD ± 0.92 years), respectively. Statistically significant differences in several self-reported baseline behaviors and attitudinal responses were found between the two groups; for example, 47.4% of previous participants compared to 29.4% of first-time participants disagreed that reading text messages only at a stop light was acceptable. Students were also asked how likely they were to intervene if a friend or family member was practicing unsafe driving behaviors; responses were similar between the two groups. The baseline behaviors and attitudes of participants regarding impaired and distracted driving were more protective among previous participants compared to first-time participants, suggesting the program results in long-term positive changes in behaviors and attitudes. The results of this secondary retrospective study may be useful for informing the implementation of future impaired and distracted driving prevention programs.