Optimization of a genomic editing system using CRISPR/Cas9-induced site-specific gene integration

The CRISPR-Cas system is an adaptive immune system found in bacteria which helps protect against the invasion of other microorganisms. This system induces double stranded breaks at precise genomic loci (1) in which repairs are initiated and insertions of a target are completed in the process. This mechanism can be used in eukaryotic cells in combination with sgRNAs (1) as a tool for genome editing. By using this CRISPR-Cas system, in addition to the “safe harbor locus,” ROSAβ26, the incorporation of a target gene into a site that is not susceptible to gene silencing effects can be achieved through few simple steps. PCR amplification of the target genes , ROSA26 and mKate2, with a sgRNA scaffold and T7 promoter followed by in vitro transcription aim to produce an RNA product. This sgRNA product can be run through a digestion with Cas9 to validate cleavage of the genomic ROSA DNA template or mKate plasmid. Osteoblast mouse cells are transfected and labeled through partial uptake by the CRISPR mechanism, by cutting in the ROSA loci and repairing with pieces of the fluorescent mKate2 plasmid. These cells were measured via flow cytometry to give a percentage of red cells. This data shows the scaffolding construct created is targeted by the Cas9 endonuclease and through homologous repair the cells will incorporate the mKate2 target gene in vitro in MC3T3 mouse cells

A comparison of hospital and nonhospital colonoscopy: Wait times, fees and guideline adherence to follow-up interval

BACKGROUND: Although colonoscopy is increasingly performed in nonhospital facilities, studies to date examining differences between colonoscopy services in hospital and nonhospital settings have been limited, in large part, to administrative databases

Addressing Motivations and Barriers to Research Involvement during Medical School among Osteopathic Medical Students in the United States

Involvement in research is regarded as a high-impact educational practice, which, for medical professionals, is associated with sharpened critical thinking and life-long learning skills, greater appreciation for evidence-based medicine, and better clinical competence scores. However, there are limited data regarding the research experience and/or interest among osteopathic medical students in the United States despite a rapidly increasing enrollment and expansion of the number of osteopathic medical schools. Thus, we administered an electronic survey examining prior research experience, interests, and perceptions about research participation during medical school to four successive classes of incoming first-year osteopathic medical students. We also performed focus groups with rising third-year osteopathic medical students around the topic of perceived barriers to and potential enablers of promoting research participation. This yielded a survey addendum where first-year osteopathic medical students provided feedback on the likelihood of specific incentives/enablers to encourage participation in research during medical school. Overall, osteopathic medical students are interested in research, view research experience as valuable, and perceive research experience as beneficial to future career development. Students perceive that the primary barrier to involvement in research is a potential negative impact upon performance in coursework. Feedback on the likelihood of specific enablers/incentives was also garnered. Our findings from a single institution may have important implications in defining the prior experiences and perceptions held by first-year osteopathic medical students. Specifically, our study indicates that research experiences intentionally designed with (1) a strong likelihood of gaining a publication, (2) financial compensation, and (3) the opportunity for short-term involvement, a flexible time commitment, and/or a dedicated time period are most likely to encourage research participation by osteopathic medical students

Addressing Motivations and Barriers to Research Involvement during Medical School among Osteopathic Medical Students in the United States

Involvement in research is regarded as a high-impact educational practice, which, for medical professionals, is associated with sharpened critical thinking and life-long learning skills, greater appreciation for evidence-based medicine, and better clinical competence scores. However, there are limited data regarding the research experience and/or interest among osteopathic medical students in the United States despite a rapidly increasing enrollment and expansion of the number of osteopathic medical schools. Thus, we administered an electronic survey examining prior research experience, interests, and perceptions about research participation during medical school to four successive classes of incoming first-year osteopathic medical students. We also performed focus groups with rising third-year osteopathic medical students around the topic of perceived barriers to and potential enablers of promoting research participation. This yielded a survey addendum where first-year osteopathic medical students provided feedback on the likelihood of specific incentives/enablers to encourage participation in research during medical school. Overall, osteopathic medical students are interested in research, view research experience as valuable, and perceive research experience as beneficial to future career development. Students perceive that the primary barrier to involvement in research is a potential negative impact upon performance in coursework. Feedback on the likelihood of specific enablers/incentives was also garnered. Our findings from a single institution may have important implications in defining the prior experiences and perceptions held by first-year osteopathic medical students. Specifically, our study indicates that research experiences intentionally designed with (1) a strong likelihood of gaining a publication, (2) financial compensation, and (3) the opportunity for short-term involvement, a flexible time commitment, and/or a dedicated time period are most likely to encourage research participation by osteopathic medical students