Objectives

To identify factors associated with Opioid Agonist Treatment (OAT) satisfaction and to determine whether these relationships are gender specific.

Methods

This study was based on data collected in a cross-sectional study among long-term opioid-dependent individuals (n = 160; 46.3% women). Participants completed the Client Satisfaction Questionnaire in reference to OAT episodes. Sociodemographic, illicit substance use, health, and addiction treatment history data were collected. Multivariable linear regression was used to determine the relationship between these variables and treatment satisfaction. To explore the potential role of gender in these identified relationships stratified multivariable models were tested. Additional open-ended questions regarding positive and negative perceptions of treatment were collected, and a thematic analysis was conducted.

Results

In the multivariable linear regression model, participants who were older, of Aboriginal ancestry, and currently receiving OAT had higher OAT satisfaction scores, whereas participants who had methadone dose preferences of 30 mg or less had lower OAT satisfaction. In stratified analyses among women, the relationship between preferred methadone dose and current OAT remained significantly associated with satisfaction. Open-ended positive and negative perceptions complemented and provided further valuable data to interpret these identified relationships.

Conclusions

To our knowledge, this is the first study to explore the potential role of gender in factors associated with OAT satisfaction. These findings provide valuable information to health care providers working in OAT settings regarding how to address women and men's OAT needs and improve treatment satisfaction.

Background

One of the bottlenecks in production of biochemicals and pharmaceuticals in Saccharomyces cerevisiae is stable and homogeneous expression of pathway genes. Integration of genes into the genome of the production organism is often a preferred option when compared to expression from episomal vectors. Existing approaches for achieving stable simultaneous genome integrations of multiple DNA fragments often result in relatively low integration efficiencies and furthermore rely on the use of selection markers.

Results

Here, we have developed a novel method, CrEdit (CRISPR/Cas9 mediated genome Editing), which utilizes targeted double strand breaks caused by CRISPR/Cas9 to significantly increase the efficiency of homologous integration in order to edit and manipulate genomic DNA. Using CrEdit, the efficiency and locus specificity of targeted genome integrations reach close to 100% for single gene integration using short homology arms down to 60 base pairs both with and without selection. This enables direct and cost efficient inclusion of homology arms in PCR primers. As a proof of concept, a non-native β-carotene pathway was reconstructed in S. cerevisiae by simultaneous integration of three pathway genes into individual intergenic genomic sites. Using longer homology arms, we demonstrate highly efficient and locus-specific genome integration even without selection with up to 84% correct clones for simultaneous integration of three gene expression cassettes.

Conclusions

The CrEdit approach enables fast and cost effective genome integration for engineering of S. cerevisiae. Since the choice of the targeting sites is flexible, CrEdit is a powerful tool for diverse genome engineering applications.