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RNA Interference and CRISPR Technologies pp 365–383Cite as

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

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2115))

Abstract

CRISPR Cas9 genome editing allows researchers to modify genes in a multitude of ways including to obtain deletions, epitope-tagged loci, and knock-in mutations. Within 6 years of its initial application, CRISPR-Cas9 genome editing has been 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.

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Authors and Affiliations

  1. Department of Biology, University of Texas – Rio Grande Valley, Edinburg, TX, USA

    Nirakar Sahoo, Victoria Cuello, Shreya Udawant, Carl Litif, Julie A. Mustard & Megan Keniry

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  1. Nirakar Sahoo
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  2. Victoria Cuello
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  3. Shreya Udawant
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  4. Carl Litif
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  5. Julie A. Mustard
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  6. Megan Keniry
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Corresponding author

Correspondence to Megan Keniry .

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Editors and Affiliations

  1. Department of Cancer Immunology, Institute for Cancer Research, Oslo University Hospital-Radiumhospitalet, Oslo, Norway

    Mouldy Sioud

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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

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  • DOI: https://doi.org/10.1007/978-1-0716-0290-4_20

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