The ORFeome Collaboration: a genome-scale human ORF-clone resource

Personalised proteome analysis by means of protein microarrays made from individual patient samples

“…In order to define the efficiency of the in situ transcription and translation process, templates were used that had been produced with common primers from 2016 full-length cDNAs, which were randomly selected by using 21 microtiter plates of a library of in total about 12,000 non-redundant, full-length and sequence-verified clones15. Prokaryotic cell lysate was used for in situ protein expression.…”

Proteome-Scale Human Interactomics

“…Indeed, it took several decades to develop robust methods for detecting PPIs at proteome-scale. These efforts paired with the availability of nearly complete “ORFeome” collections of “readyto-be-expressed” human open reading frames (ORFs) [2] laid the necessary foundation to enable substantial progress in mapping the human PPI interactome (hereafter referred to as “interactome” for simplicity). In the span of a couple of years, four groundbreaking human interactome maps have been published (Figure 1), each utilizing different methodologies and thus capturing different aspects of the human interactome [3–6].…”

“…For example, Y2H-based PPI mapping transitioned from using cDNA libraries to using sequencevalidated and arrayable sets of ORF clones during screening [22]. These sets have grown into near-complete ‘human ORFeome collections’, resources of protein-encoding ORF clones with a representative protein for almost every human gene [2]. Furthermore, rigorous quality control measures were implemented that eliminated spontaneous autoactivation of DNA binding-fusion proteins (i.e., activation of the reporter gene in the absence of the activation domain-containing fusion construct) [7, 23].…”

Comprehensive Analyses of Tissue-Specific Networks with Implications to Psychiatric Diseases

“…These and similar studies identify thousands of PPIs by systematically testing large number of human proteins against the entire human ORFeome [27] using consistent experimental procedures with follow-up pair-wise verification by independent methods [28]. Although such selection strategy may decrease the number of PPIs that could be used for network construction, compared to those available from large public databases such as STRING [29] or InWeb [30], selection of unbiased PPIs would likely increase confidence and reliability of the resulting networks.…”