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CRISPR-Cas9 is revolutionizing how we conduct scientific research, treat disease, and develop new crops. The widespread impact of this genome-editing technology makes it critical for undergraduate students to understand and engage with this new tool. In this article, we describe a multi-week lab module that teaches undergraduates how to design CRISPR-Cas9 constructs and screen for CRISPR-modified genotypes. The module is conducted through the lens of independent research; students conduct a genotype screen for novel knockout mutations. In our module, students screen Zea mays (maize) seedlings for mutations in the MAD2 gene, which assists our ongoing investigation of meiotic chromosome segregation. This module can be adapted to knockout any gene in any organism, and thus align with an instructor’s research program. Engaging in original research helps undergraduate students develop independence and initiative in the lab as well as the molecular techniques of CRISPR-Cas9.
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Beyond editing, CRISPR/Cas9 for protein localization: an educational primer for use with “A dCas9-based system identifies a central role for Ctf19 in kinetochore-derived suppression of meiotic recombination”
Abstract CRISPR/Cas9 has dramatically changed how we conduct genetic research, providing a tool for precise sequence editing. However, new applications of CRISPR/Cas9 have emerged that do not involve nuclease activity. In the accompanying article “A dCas9-based system identifies a central role for Ctf19 in kinetochore-derived suppression of meiotic recombination,” Kuhl et al. utilize a catalytically dead Cas9 to localize proteins at specific genomic locations. The authors seek to understand the role of kinetochore proteins in the suppression of meiotic recombination, a phenomenon that has been observed in centromere regions. By harnessing the power of CRISPR/Cas9 to bind specific genomic sequences, Kuhl et al. localized individual kinetochore proteins to areas of high meiotic recombination and assessed their role in suppression. This primer article provides undergraduate students with background information on chromosomes, meiosis, recombination and CRISPR/Cas9 to support their reading of the Kuhl et al. study. This primer is intended to help students and instructors navigate the study’s experimental design, interpret the results, and appreciate the broader scope of meiotic recombination and CRISPR/Cas9. Questions are included to facilitate discussion of the study.
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- Award ID(s):
- 1841696
- PAR ID:
- 10370923
- Publisher / Repository:
- Oxford University Press
- Date Published:
- Journal Name:
- Genetics
- Volume:
- 222
- Issue:
- 1
- ISSN:
- 1943-2631
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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