Programmable promoter editing for precise control of transgene expression.

Subtle changes in gene expression direct cells to distinct cellular states. Identifying and controlling dose-dependent transgenes requires tools for precisely titrating expression. To this end, we developed a framework called DIAL for building editable promoters that allows for fine-scale, heritable changes in transgene expression. Using DIAL, we increase expression by recombinase-mediated excision of spacers between the binding sites of a synthetic zinc-finger transcription factor and the core promoter. By nesting varying numbers and lengths of spacers, DIAL generates a tunable range of unimodal setpoints from a single promoter construct. Through small-molecule control of transcription factors and recombinases, DIAL supports temporally defined, user-guided control of transgene expression. Integration of DIAL promoters into lentivirus allows for efficient delivery to primary cells. As promoter editing generates stable states, DIAL setpoints are heritable, facilitating mapping of transgene levels to phenotypes. The highly modular and extensible DIAL framework opens up new opportunities for screening and tailoring transgene expression to regulate gene and cell-based therapies.

Proliferation history and transcription factor levels drive direct conversion.

The sparse and stochastic nature of reprogramming has obscured our understanding of how transcription factors drive cells to new identities. To overcome this limit, we developed a compact, portable reprogramming system that increases direct conversion of fibroblasts to motor neurons by two orders of magnitude. We show that subpopulations with different reprogramming potentials are distinguishable by proliferation history. By controlling for proliferation history and titrating each transcription factor, we find that conversion correlates with levels of the pioneer transcription factor Ngn2, whereas conversion shows a biphasic response to Lhx3. Increasing the proliferation rate of adult human fibroblasts generates morphologically mature, induced motor neurons at high rates. Using compact, optimized, polycistronic cassettes, we generate motor neurons that graft with the murine central nervous system, demonstrating the potential for in vivo therapies.

Friendships and Self-Determination Among Students Who Are Deaf or Hard of Hearing.

Friendships and self-determination are two factors that positively influence success in school and adult life. Examining the relationship between these two constructs among a sample of students who were deaf or hard of hearing (DHH), the researchers used the Self-Determination Inventory: Student Report (Shogren, Wehmeyer, Burke, & Palmer, 2017) to measure the level of self-determination of 29 students ages 13-17 years who were DHH. The researchers then used two self-report measures to probe students' quantity and quality of friendships, and conducted an analysis to determine whether the quality and quantity of friendships correlated with the self-determination of the students. Significant correlations were found between self-determination and both the quality and quantity of friendships among the study sample (p < .01). The findings support those of previous studies indicating that autonomy and social networks are related.

Job Satisfaction of Teachers of Students who are Deaf or Hard of Hearing.

The pressure that educators are experiencing to educate more students, with more challenges, to higher levels of learning than any time in the past is significantly changing their working conditions. The purpose of this study was to examine the perceptions of a national sample of teachers of students who are deaf or hard of hearing to ascertain their overall job satisfaction as well as to identify the specific factors that positively and negatively affect their ability to do their jobs well. In addition, responses of different subsets of teachers (e.g., itinerant, elementary, and secondary) were compared. Responses from 495 deaf educators are reported. Collectively, 89% of participants stated that they were satisfied to very satisfied with their overall job. Specific aspects of the job that respondents indicated that they were most satisfied or most dissatisfied with are presented and suggestions for addressing some of the identified challenges as well as recommendations for future research are provided.