Peer-reviewedCC BY-NC-ND 4.0STAR Protocols
Protocol for eVLP-mediated CRISPR-Cas9 and prime editing in pancreatic cancer cell lines
7 affiliations
- 1Liverpool Experimental Cancer Medicine Centre, University of Liverpool, Liverpool L69 3GE, UK.
- 2Department of Biochemistry and Molecular Biology, Faculty of Biological Sciences, Complutense University of Madrid, Madrid, Spain; San Carlos Health Research Institute (IdISSC), 28040 Madrid, Spain.
- 3Department of Medical Oncology, Cancer Center Amsterdam, Amsterdam University Medical Center, Vrije Universiteit, Amsterdam, the Netherlands; Cancer Pharmacology Lab, Fondazione Pisana per La Scienza, Pisa, San Giuliano Terme, Italy.
- 4School of Medicine, Pharmacy and Biomedical Sciences, University of Portsmouth, Portsmouth, UK.
- 5Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, Complutense University of Madrid, Madrid, Spain; San Carlos Health Research Institute (IdISSC), 28040 Madrid, Spain.
- 6Liverpool Experimental Cancer Medicine Centre, University of Liverpool, Liverpool L69 3GE, UK. Electronic address: [email protected].
- 7Liverpool Experimental Cancer Medicine Centre, University of Liverpool, Liverpool L69 3GE, UK. Electronic address: [email protected].
Abstract
Pancreatic ductal adenocarcinoma (PDAC) cell lines resist transfection, limiting gene-editing experiments. Here, we present a protocol for delivering CRISPR-Cas9 or prime editors into PDAC cell lines using retrovirus-derived engineered virus-like particles (eVLPs). We describe steps for generating eVLPs, packaging CRISPR-Cas9 ribonucleoproteins or prime editors with engineered prime-editing guide RNAs (epegRNAs), and transducing human PDAC cells. We use the HEK3 locus and NQO1 (NAD(P)H quinone dehydrogenase 1) exon 6 as example edits and controls, respectively. This protocol adapts readily to other genomic sites.
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BibTeX
@article{timminsyoung2026protocol,
title = {{Protocol for eVLP-mediated CRISPR-Cas9 and prime editing in pancreatic cancer cell lines}},
author = {Chloe Timmins-Young and Marco Cordani and Elisa Giovannetti and Despoina Aslanoglou and Alvaro Gutierrez and Eithne Costello and Mark Glenn},
journal = {STAR Protocols},
year = {2026},
month = oct,
doi = {10.1016/j.xpro.2026.104904},
url = {https://doi.org/10.1016/j.xpro.2026.104904},
note = {CC BY-NC-ND 4.0},
}
RIS
TY - JOUR
TI - Protocol for eVLP-mediated CRISPR-Cas9 and prime editing in pancreatic cancer cell lines
AU - Chloe Timmins-Young
AU - Marco Cordani
AU - Elisa Giovannetti
AU - Despoina Aslanoglou
AU - Alvaro Gutierrez
AU - Eithne Costello
AU - Mark Glenn
PY - 2026
DA - 2026/10/17
JO - STAR Protocols
DO - 10.1016/j.xpro.2026.104904
UR - https://doi.org/10.1016/j.xpro.2026.104904
AB - Pancreatic ductal adenocarcinoma (PDAC) cell lines resist transfection, limiting gene-editing experiments. Here, we present a protocol for delivering CRISPR-Cas9 or prime editors into PDAC cell lines using retrovirus-derived engineered virus-like particles (eVLPs). We describe steps for generating eVLPs, packaging CRISPR-Cas9 ribonucleoproteins or prime editors with engineered prime-editing guide RNAs (epegRNAs), and transducing human PDAC cells. We use the HEK3 locus and NQO1 (NAD(P)H quinone dehydrogenase 1) exon 6 as example edits and controls, respectively. This protocol adapts readily to other genomic sites.
SN - 2666-1667
ER -