An Undergraduate Laboratory Experience Using CRISPR-cas9 Technology to Deactivate Green Fluorescent Protein Expression in "Escherichia coli"

Autor/inn/en	Pieczynski, Jay N.; Deets, Amber; McDuffee, Alexis; Lynn Kee, H.
Titel	An Undergraduate Laboratory Experience Using CRISPR-cas9 Technology to Deactivate Green Fluorescent Protein Expression in "Escherichia coli"
Quelle	In: Biochemistry and Molecular Biology Education, 47 (2019) 2, S.145-155 (11 Seiten)Infoseite zur Zeitschrift PDF als Volltext Verfügbarkeit
Zusatzinformation	ORCID (Lynn Kee, H.)
Sprache	englisch
Dokumenttyp	gedruckt; online; Zeitschriftenaufsatz
ISSN	1470-8175
DOI	10.1002/bmb.21206
Schlagwörter	Genetics; Science Laboratories; Active Learning; Hands on Science; College Science; Undergraduate Students; Science Instruction; Biotechnology; Technology Uses in Education; Instructional Effectiveness + Suchen Sie Ihr Suchwort? Humangenetik; Aktives Lernen; Teaching of science; Science education; Natural sciences Lessons; Naturwissenschaftlicher Unterricht; Biotechnologie; Technology enhanced learning; Technology aided learning; Technologieunterstütztes Lernen; Unterrichtserfolg
Abstract	Undergraduates learn that gene editing in diverse organisms is now possible. How targeted manipulation of genes and genomes is utilized in basic science and biomedicine to address biological questions is challenging for undergraduates to conceptualize. Thus, we developed a lab experience that would allow students to be actively engaged in the full process of design, implementation of a gene editing strategy, and interpretation of results within an 8-week lab period of a Genetics course. The laboratory experience combines two transformative biotechnology tools; the utilization of green fluorescent protein (GFP) as a diagnostic marker of gene expression and the fundamentals and specificity of Clustered Regularly Interspaced Short Palindromic Repeats-cas9 (CRISPR-cas9) gene editing in bacterial cells. The students designed and constructed plasmids that express single guide RNA targeted to GFP, expressed the sgRNA and cas9 in bacteria cells, and successfully deactivated GFP gene expression in the bacterial cells with their designed CRISPR-cas9 tools. Student assessment revealed most students achieved student learning objectives. We conclude this lab experience is an effective and accessible method for engaging students in the scientific practices, knowledge and challenges revolving targeted CRISPR-cas9 gene manipulation. (As Provided).
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Erfasst von	ERIC (Education Resources Information Center), Washington, DC
Update	2020/1/01