The Genetic Code Kit: An Open-Source Cell-Free Platform for Biochemical and Biotechnology Education.

Teaching the processes of transcription and translation is challenging due to the intangibility of these concepts and a lack of instructional, laboratory-based, active learning modules. Harnessing the genetic code in vitro with cell-free protein synthesis (CFPS) provides an open platform that allows for the direct manipulation of reaction conditions and biological machinery to enable inquiry-based learning. Here, we report our efforts to transform the research-based CFPS biotechnology into a hands-on module called the "Genetic Code Kit" for implementation into teaching laboratories. The Genetic Code Kit includes all reagents necessary for CFPS, as well as a laboratory manual, student worksheet, and augmented reality activity. This module allows students to actively explore transcription and translation while gaining exposure to an emerging research technology. In our testing of this module, undergraduate students who used the Genetic Code Kit in a teaching laboratory showed significant score increases on transcription and translation questions in a post-lab questionnaire compared with students who did not participate in the activity. Students also demonstrated an increase in self-reported confidence in laboratory methods and comfort with CFPS, indicating that this module helps prepare students for careers in laboratory research. Importantly, the Genetic Code Kit can accommodate a variety of learning objectives beyond transcription and translation and enables hypothesis-driven science. This opens the possibility of developing Course-Based Undergraduate Research Experiences (CUREs) based on the Genetic Code Kit, as well as supporting next-generation science standards in 8-12th grade science courses.

Unlocking Applications of Cell-Free Biotechnology through Enhanced Shelf Life and Productivity of E. coli Extracts.

Cell-free protein synthesis (CFPS) is a platform biotechnology that enables a breadth of applications. However, field applications remain limited due to the poor shelf-stability of aqueous cell extracts required for CFPS. Lyophilization of E. coli extracts improves shelf life but remains insufficient for extended storage at room temperature. To address this limitation, we mapped the chemical space of ten low-cost additives with four distinct mechanisms of action in a combinatorial manner to identify formulations capable of stabilizing lyophilized cell extract. We report three key findings: (1) unique additive formulations that maintain full productivity of cell extracts stored at 4 °C and 23 °C; (2) additive formulations that enhance extract productivity by nearly 2-fold; (3) a machine learning algorithm that provides predictive capacity for the stabilizing effects of additive formulations that were not tested experimentally. These findings provide a simple and low-cost advance toward making CFPS field-ready and cost-competitive for biomanufacturing.

A Meta-analytic Review of the Five Minute Speech Sample as a Measure of Family Emotional Climate for Youth: Relations with Internalizing and Externalizing Symptomatology.

The Five Minute Speech Sample's (FMSS) measure of parental expressed emotion (EE), defined as criticism (CRIT) and emotional overinvolvement (EOI), has been increasingly used to measure family emotional climate in relation to youth psychopathological development. As CRIT and EOI were defined based on adults, a meta-analysis and systematic review was conducted to analyze the presence and strength of an effect among maternal CRIT and EOI with youth internalizing and externalizing problems. A random effects model was used to analyze the 42 studies on families of youth (aged 1.5 to 19). There was a small, significant relation among maternal CRIT with youth internalizing and externalizing problems and among EOI with youth internalizing problems. EOI was not significantly related to externalizing problems. The current study suggests that the FMSS measure of CRIT is a more robust correlate of youth internalizing and externalizing symptoms than EOI, but EOI does relate to internalizing behaviors. Few moderators emerged, highlighting a continued need to identify factors accounting for heterogeneity. The current results suggest that the FMSS measure of CRIT may be a valuable measure of the family emotional climate in families of youth, but care should be taken when including analyses on EOI.