Homo sapiens
Protocols: >1000-fold coverage of mutagenized cells (Brunello library) was infected with PeV-A1 or PeV-A2 at MOI 0.1. Resistant cell colonies were harvested. Uninfected starting populations of mutagenized cells were used as the unselected reference. Total genomic DNA from both virus-resistant and uninfected cells was respectively extracted using QIAamp DNA Mini Kit (Qiagen). A first round of PCR was used to amplify the guide RNA sequences encoded in the gDNA, followed by a second round of PCR to add the barcodes/adapters for amplicon sequencing. 2% agarose gels and a QIAquick gel extraction kit were used to purify the amplicons.
disease
Intestinal Type Adenocarcinoma
organism
Homo sapiens
organism
Homo sapiens
collection date
not collected
scientific_name
Homo sapiens
common name
human
organism part
colon
genotype
lentiCas9-Blast; lentiGuide-Puro; SLC35A1/EXTL3 knockout, PeV-A1 infection
isolate
not applicable
cell_line
HT-29
cell_type
epithelial cell
geographic location (country and/or sea)
not collected
infect
PeV-A1