CRISPR Cas9 Genome Editing in Human Cell Lines with DONOR Vector Made by Gibson Assembly

Authors

Nirakar Sahoo, The University of Texas Rio Grande Valley
Victoria Cuello, The University of Texas Rio Grande Valley
Shreya Udawant, The University of Texas Rio Grande Valley
Carl Litif, The University of Texas Rio Grande Valley
Julie A. Mustard, The University of Texas Rio Grande Valley
Megan Keniry, The University of Texas Rio Grande Valley

Document Type

Book

Publication Date

2-2020

Abstract

CRISPR Cas9 genome editing allows researchers to modify genesin a multitude of ways including to obtain deletions, epitope-tagged loci, and knock-in mutations. Within six years of its initial application, CRISPR Cas9 genome editing has become widely employed, but disadvantages to this method, such as low modification efficiencies and off-target effects,need careful consideration. Obtaining custom donor vectors can also be expensive and time consuming. This chapter details strategies to overcome barriers to CRISPR Cas9 genome editing as well as recent developments in employing this technique.

Comments

© Springer Science+Business Media, LLC, part of Springer Nature 2020. Original published version available at https://doi.org/10.1007/978-1-0716-0290-4_20

Recommended Citation

Sahoo N., Cuello V., Udawant S., Litif C., Mustard J.A., Keniry M. (2020) CRISPR-Cas9 Genome Editing in Human Cell Lines with Donor Vector Made by Gibson Assembly. In: Sioud M. (eds) RNA Interference and CRISPR Technologies. Methods in Molecular Biology, vol 2115. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0290-4_20

Publication Title

Methods in Molecular Biology

DOI

10.1007/978-1-0716-0290-4_20