Fire Burn and Cauldron Bubble: What is in your Genome Editing Brew?

Published

October 28, 2022

Author(s)

Simona Patange, Samantha Maragh

Abstract

Genome editing is a rapidly evolving biotechnology with the potential to transform many sectors of industry such as agriculture, biomanufacturing, and medicine. This technology is enabled by an ever-growing portfolio of biomolecular reagents that span the central dogma, from DNA to RNA to protein. In this paper, we draw from our unique perspective as the National Metrology Institute of the United States to bring attention to the importance of understanding and reporting genome editing formulations accurately and promoting concepts to verify successful delivery into cells. Achieving the correct understanding may be hindered by the way units, quantities, and stoichiometries are reported in the field. We highlight the variability in how editing formulations are reported in the literature and examine how a reference molecule could be used to verify the delivery of a reagent into cells. We provide recommendations on how more accurate reporting of editing formulations and more careful verification of the steps in an editing experiment can help set baseline expectations of reagent performance, toward the aim of enabling genome editing studies to be more reproducible. We conclude with a future outlook on technologies that can further our control and enable our understanding of genome editing outcomes at the single-cell level.

Citation

ACS Biochemistry

Pub Type

Journals

Download Paper

https://doi.org/10.1021/acs.biochem.2c00431

Local Download

Keywords

CRISPR, Cas9, gRNA, gene editing, genome editing, single-cell, biomolecule, biochemistry, fluorescence

Citation

Patange, S. and Maragh, S. (2022), Fire Burn and Cauldron Bubble: What is in your Genome Editing Brew?, ACS Biochemistry, [online], https://doi.org/10.1021/acs.biochem.2c00431, https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=935227 (Accessed November 23, 2024)