title: Improved delivery of Cas9 protein/gRNA complexes using lipofectamine CRISPRMAX.
abstract: OBJECTIVE
  To identify the best lipid nanoparticles for delivery of purified Cas9 protein and gRNA complexes (Cas9 RNPs) into mammalian cells and to establish the optimal conditions for transfection.
  
  
  RESULTS
  Using a systematic approach, we screened 60 transfection reagents using six commonly-used mammalian cell lines and identified a novel transfection reagent (named Lipofectamine CRISPRMAX). Based on statistical analysis, the genome modification efficiencies in Lipofectamine CRISPRMAX-transfected cell lines were 40 or 15 % higher than those in Lipofectamine 3000 or RNAiMAX-transfected cell lines, respectively. Upon optimization of transfection conditions, we observed 85, 75 or 55 % genome editing efficiencies in HEK293FT cells, mouse ES cells, or human iPSCs, respectively. Furthermore, we were able to co-deliver donor DNA with Cas9 RNPs into a disrupted EmGFP stable cell line, resulting in the generation of up to 17 % EmGFP-positive cells.
  
  
  CONCLUSIONS
  Lipofectamine CRISPRMAX was characterized as the best lipid nanoparticles for the delivery of Cas9 RNPs into a variety of mammalian cell lines, including mouse ES cells and iPSCs.
authors.affiliation: Synthetic Biology Department, Thermo Fisher Scientific, 5781 Van Allen Way, Carlsbad, CA, 92008, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Yu
authors.affiliation: Synthetic Biology Department, Thermo Fisher Scientific, 5781 Van Allen Way, Carlsbad, CA, 92008, USA. xiquan.liang@thermofisher.com.
authors.forename: Xiquan
authors.identifier: http://orcid.org/0000-0003-1027-3427
authors.initials: X
authors.lastname: Liang
authors.affiliation: Synthetic Biology Department, Thermo Fisher Scientific, 5781 Van Allen Way, Carlsbad, CA, 92008, USA.
authors.forename: Huimin
authors.initials: H
authors.lastname: Xie
authors.affiliation: Synthetic Biology Department, Thermo Fisher Scientific, 5781 Van Allen Way, Carlsbad, CA, 92008, USA.
authors.forename: Shantanu
authors.initials: S
authors.lastname: Kumar
authors.affiliation: Synthetic Biology Department, Thermo Fisher Scientific, 5781 Van Allen Way, Carlsbad, CA, 92008, USA.
authors.forename: Namritha
authors.initials: N
authors.lastname: Ravinder
authors.affiliation: Synthetic Biology Department, Thermo Fisher Scientific, 5781 Van Allen Way, Carlsbad, CA, 92008, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Potter
authors.affiliation: Synthetic Biology Department, Thermo Fisher Scientific, 5781 Van Allen Way, Carlsbad, CA, 92008, USA.
authors.forename: Xavier
authors.initials: X
authors.lastname: de Mollerat du Jeu
authors.affiliation: Synthetic Biology Department, Thermo Fisher Scientific, 5781 Van Allen Way, Carlsbad, CA, 92008, USA.
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Chesnut
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D002460:Cell Line; D018274:Electroporation; D000072669:Gene Editing / Q000379:methods; D018390:Gene Targeting / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells; D008055:Lipids / Q000633:toxicity; D051379:Mice; D014162:Transfection / Q000379:methods*
chemical_list: D008055:Lipids; C086724:Lipofectamine; D049452:Green Fluorescent Proteins
keywords: CRISPR; CRISPRMAX; Cas9 protein; Cell engineering; Genome editing; Homologous recombination