Open AccessCellular thermal shift analysis for interrogation of CRISPR-assisted proteomic changes
Author(s) -
Nam-Gu Her,
Ivan Babić,
Venkata Mahidhar Yenugonda,
Santosh Kesari,
Elmar Nurmemmedov
Publication year - 2020
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2019-0100
Subject(s) - crispr , computational biology , proteomics , mutagenesis , biology , functional genomics , cas9 , genomics , genetics , gene , genome , mutation
CRISPR–Cas9 has proven to be a versatile tool for the discovery of essential genetic elements involved in various disease states. CRISPR-assisted dense mutagenesis focused on therapeutically challenging protein complexes allows us to systematically perturb protein-coding sequences in situ and correlate them with functional readouts. Such perturbations can mimic targeting by therapeutics and serve as a foundation for the discovery of highly specific modulators. However, translation of such genomics data has been challenging due to the missing link for proteomics under the physiological state of the cell. We present a method based on cellular thermal shift assays to easily interrogate proteomic shifts generated by CRISPR-assisted dense mutagenesis, as well as a case focused on NuRD epigenetic complex.