Pageblue solution

Unmodified crRNA, 5’ Amine-crRNA, 5’ Azide-crRNA, 5’ Thiol-crRNA, 3’ Amine-crRNA, 5’ Amine-Donor, 3’ Amine-Donor, 5’ Azide-Donor, and various DNA sequences were purchased from Integrated DNA Technology (IDT). Phusion High-fidelity DNA Polymerase was purchased from NEB (Ipswich, MA). The Megascript T7 kit, the Megaclear kit, the PageBlue solution, the propidium iodide, and the PureLink genomic DNA kit were purchased from Thermo Fisher (Waltham, MA). Mini-PROTEAN TGX Gels (4–20%) were purchased from Bio-Rad (Hercules, CA). 4-(2-hydroxyethyl) piperazine-1-ethanesulfonate (HEPES) were purchased from Mandel Scientific (Guelph, ON). Sodium silicate was purchased from Sigma Aldrich (St. Louis, MO). Matrigel was purchased from BD Biosciences (San Jose, CA). DMEM media, non-essential amino acids, penicillin-streptomycin, DPBS and 0.05% trypsin were purchased from Life Technologies (Carlsbad, CA). EMD Millipore Amicon Ultra-4 100 kDa and 300 kDa were purchased from Millipore (Germany). Streptococcus pyogenes Cas9 (spCas9) and Acidaminococcus sp. Cpf1 (AsCpf1) were purchased from the QB3 Macrolab from UC Berkeley. PAsp(DET) polymer was a generous gift from Dr. Kataoka’s group (Miyata et al., 2008 (link); Kim et al., 2010 (link), 2014 (link)).

SpCas9 and AsCpf1 were purchased from the MacroLab in UC Berkeley. Proteins were stored in 50 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) at pH 7.5 with 300 mM NaCl, 10% glycerol, and 100 μM tris(2-carboxyethyl) phosphine at −80 °C. PAsp(DET) polymer was synthesized using the following literature references^{46 (link),60 (link),61 (link)}. Phusion High-fidelity DNA Polymerase was purchased from NEB (Ipswich, MA, USA). The MEGAscript T7 kit, the MEGAclear kit, the PageBlue solution, the propidium iodide, and the PureLink genomic DNA kit were purchased from Thermo Fisher (Waltham, MA, USA). Mini-PROTEAN TGX (Tris-Glycine eXtended) gels (4–20%) were purchased from Bio-Rad (Hercules, CA, USA). Dulbecco’s modified Eagle's medium (DMEM), non-essential amino acids, penicillin–streptomycin, Dulbecco's phosphate-buffered saline (PBS), and 0.05% trypsin were purchased from Life Technologies (Carlsbad, CA, USA).