Table 5.
Advantages and disadvantages of Y2H and Y1H assays
| Y2H advantages | Y2H disadvantages | Y1H advantages | Y1H disadvantages |
|---|---|---|---|
| Condition-independent; can use low-abundance proteins that are difficult to detect biochemically | Not done in endogenous context; may retrieve irrelevant PPIs (biological false positives) | Condition-independent; can identify low-abundance and tissue-restricted TFs that are difficult to detect by ChIP | Not done in endogenous context; may retrieve irrelevant PDIs (biological false positives) |
| Can detect binary PPIs | Some types of proteins cannot be effectively used, such as membrane proteins | Can identify multiple TFs that bind a DNA fragment of interest (gene centered) | Not (yet) adapted for use with TF heterodimers |
| Can detect transient PPIs | PPIs that depend on posttranslational modifications are usually not detected | Can detect weak PDIs | PDIs that depend on posttranslational modifications are usually not detected. |
| Can be used to delineate interaction domains | Can be used to delineate and refine TF binding sites | ||
| Can be used to identify interaction-defective alleles | Can identify novel putative TFs |
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Y2H, yeast two-hybrid; Y1H, yeast one-hybrid; PPIs, protein–protein interactions; TF, transcription factor; ChIP, chromatin immunoprecipitation; PDIs, protein–DNA interactions.
