Use of CRISPR-Cas9 Machinery to Generate Potential IFIT3 Knockout in A549 Cells
Faculty Sponsor(s)
Jennifer Grier
Location
Jereld R. Nicholson Library: Grand Avenue
Subject Area
Biochemistry
Description
CRISPR-Cas9 was discovered to be an effective and cheap method of generating mutations in cell lines, requiring materials that would be commonly found in an undergraduate educational lab. CRISPR-Cas9 uses guide RNAs (sgRNA) complementary to a target DNA sequence to make site-specific cuts in the DNA. The sgRNAs were generated and the resulting oligonucleotides inserted into plasmids. The sequences of the plasmids containing sgRNAs were confirmed. The plasmids were then transfected into A549 lung cells. In this work, CRISPR-Cas9 is used to generate potential IFIT3 knockouts in a human cell line, with the intention of researching impacts to functionality of the IFIT3 protein.
Recommended Citation
Bishop, Lauren; Wood, Victoria J.; Keyes, Emma; and Perez-Cervantes, Leslie, "Use of CRISPR-Cas9 Machinery to Generate Potential IFIT3 Knockout in A549 Cells" (2017). Linfield University Student Symposium: A Celebration of Scholarship and Creative Achievement. Event. Submission 24.
https://digitalcommons.linfield.edu/symposium/2017/all/24
Use of CRISPR-Cas9 Machinery to Generate Potential IFIT3 Knockout in A549 Cells
Jereld R. Nicholson Library: Grand Avenue
CRISPR-Cas9 was discovered to be an effective and cheap method of generating mutations in cell lines, requiring materials that would be commonly found in an undergraduate educational lab. CRISPR-Cas9 uses guide RNAs (sgRNA) complementary to a target DNA sequence to make site-specific cuts in the DNA. The sgRNAs were generated and the resulting oligonucleotides inserted into plasmids. The sequences of the plasmids containing sgRNAs were confirmed. The plasmids were then transfected into A549 lung cells. In this work, CRISPR-Cas9 is used to generate potential IFIT3 knockouts in a human cell line, with the intention of researching impacts to functionality of the IFIT3 protein.
