Journal of Stem Cell and Regenerative BiologyJournal of Stem Cell and Regenerative BiologyJournal of Stem Cell and Regenerative BiologyJournal of Stem Cell and Regenerative Biology2471-0598Ommega Online PublishersNew Jersey, USA63010.15436/2471-0598.15.004Research ArticleGene Editing in Human Pluripotent Stem Cells: Choosing the Correct PathGene Editing in Human Pluripotent Stem Cells: Choosing the Correct PathAmar M. Singh 1Transposagen Biopharmaceuticals Kentucky USA 2Hera BioLabs Kentucky USA Editor* E-mail: asingh@transposagenbio.com
The authors have declared that no competing interests exist.
20150511201511JSCRB-15-MRW-63023102015301020152015Creative Commons Attribution LicenseThis is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. emsp emsp The recent emergence of targeted nucleases has opened up new opportunities for performing genetic modifications with human pluripotent stem cells hPSCs These modifications can range from the creation of a routine knock-out to the more challenging single point-mutation For both the new and established user deciding on the best approach for the specific modification of interest can be an arduous task as new and improved technologies are rapidly and continuously being developed The choices between the reagents and methodologies depend entirely on the end-goal of the experiments and the locus to be modified Investigators need to decide on the best nuclease to use for each experiment from among Zinc-Finger Nucleases ZFNs Transcription Activator-Like Effector Nucleases TALENs and Clustered Regularly Interspaced Short Palindromic Repeats CRISPR Cas9 that would result in the highest likelihood of success with the fewest pitfalls Furthermore there have been significant improvements over the first-generation nucleases such as the development of the dimeric CRISPR RNA-guided Fok1 nucleases RFNs marketed as NextGENTM CRISPR that reduces the ldquo off-target rdquo mutation rate providing further options for investigators Should researchers need to perform a point mutation then considerations must be made between using single-stranded oligo-deoxynucleotides ssODN as the donor for homology-directed repair or utilizing a selection cassette within a donor vector in combination with an excision-only piggyBac trade transposase to leave a seamless edit In this review we will provide a general overview of the current technologies along with methodologies for generating point mutations while considering both their pros and cons 10