Matches in SemOpenAlex for { <https://semopenalex.org/work/W4288045147> ?p ?o ?g. }
- W4288045147 endingPage "110439" @default.
- W4288045147 startingPage "110439" @default.
- W4288045147 abstract "High-throughput assay systems have had a large impact on understanding the mechanisms of basic cell functions. However, high-throughput assays that directly assess molecular functions are limited. Herein, we describe the “GigaAssay”, a modular high-throughput one-pot assay system for measuring molecular functions of thousands of genetic variants at once. In this system, each cell was infected with one virus from a library encoding thousands of Tat mutant proteins, with each viral particle encoding a random unique molecular identifier (UMI). We demonstrate proof of concept by measuring transcription of a GFP reporter in an engineered reporter cell line driven by binding of the HIV Tat transcription factor to the HIV long terminal repeat. Infected cells were flow-sorted into 3 bins based on their GFP fluorescence readout. The transcriptional activity of each Tat mutant was calculated from the ratio of signals from each bin. The use of UMIs in the GigaAssay produced a high average accuracy (95%) and positive predictive value (98%) determined by comparison to literature benchmark data, known C-terminal truncations, and blinded independent mutant tests. Including the substitution tolerance with structure/function analysis shows restricted substitution types spatially concentrated in the Cys-rich region. Tat has abundant intragenic epistasis (10%) when single and double mutants are compared." @default.
- W4288045147 created "2022-07-27" @default.
- W4288045147 creator A5005315474 @default.
- W4288045147 creator A5007493680 @default.
- W4288045147 creator A5013237430 @default.
- W4288045147 creator A5014890138 @default.
- W4288045147 creator A5016993243 @default.
- W4288045147 creator A5017100036 @default.
- W4288045147 creator A5020526599 @default.
- W4288045147 creator A5020964443 @default.
- W4288045147 creator A5040067165 @default.
- W4288045147 creator A5045121596 @default.
- W4288045147 creator A5047078954 @default.
- W4288045147 creator A5048904735 @default.
- W4288045147 creator A5048914091 @default.
- W4288045147 creator A5049861618 @default.
- W4288045147 creator A5062855833 @default.
- W4288045147 creator A5080164890 @default.
- W4288045147 date "2022-07-01" @default.
- W4288045147 modified "2023-09-27" @default.
- W4288045147 title "GigaAssay – An adaptable high-throughput saturation mutagenesis assay platform" @default.
- W4288045147 cites W1575542444 @default.
- W4288045147 cites W1579727814 @default.
- W4288045147 cites W1591626938 @default.
- W4288045147 cites W1896714280 @default.
- W4288045147 cites W1970939786 @default.
- W4288045147 cites W1974747011 @default.
- W4288045147 cites W1976928071 @default.
- W4288045147 cites W1977186059 @default.
- W4288045147 cites W1979753840 @default.
- W4288045147 cites W1990242778 @default.
- W4288045147 cites W1991142954 @default.
- W4288045147 cites W1991649041 @default.
- W4288045147 cites W2014672632 @default.
- W4288045147 cites W2025886100 @default.
- W4288045147 cites W2036897871 @default.
- W4288045147 cites W2051236857 @default.
- W4288045147 cites W2056664929 @default.
- W4288045147 cites W2063579599 @default.
- W4288045147 cites W2063769322 @default.
- W4288045147 cites W2066958539 @default.
- W4288045147 cites W2068211039 @default.
- W4288045147 cites W2070217906 @default.
- W4288045147 cites W2070360032 @default.
- W4288045147 cites W2083223059 @default.
- W4288045147 cites W2084976646 @default.
- W4288045147 cites W2086584161 @default.
- W4288045147 cites W2094058089 @default.
- W4288045147 cites W2116699248 @default.
- W4288045147 cites W2120231504 @default.
- W4288045147 cites W2127284613 @default.
- W4288045147 cites W2127986230 @default.
- W4288045147 cites W2131271579 @default.
- W4288045147 cites W2133073150 @default.
- W4288045147 cites W2138834926 @default.
- W4288045147 cites W2140437023 @default.
- W4288045147 cites W2146849562 @default.
- W4288045147 cites W2157107905 @default.
- W4288045147 cites W2160557659 @default.
- W4288045147 cites W2174114678 @default.
- W4288045147 cites W2229824029 @default.
- W4288045147 cites W2263875137 @default.
- W4288045147 cites W2264578395 @default.
- W4288045147 cites W2329036584 @default.
- W4288045147 cites W2409176786 @default.
- W4288045147 cites W2530804451 @default.
- W4288045147 cites W2562684256 @default.
- W4288045147 cites W2582610345 @default.
- W4288045147 cites W2611239951 @default.
- W4288045147 cites W2749679400 @default.
- W4288045147 cites W2774216375 @default.
- W4288045147 cites W2783321959 @default.
- W4288045147 cites W2793570603 @default.
- W4288045147 cites W2802451173 @default.
- W4288045147 cites W2806890793 @default.
- W4288045147 cites W2868860358 @default.
- W4288045147 cites W2884441543 @default.
- W4288045147 cites W2889874867 @default.
- W4288045147 cites W2902929398 @default.
- W4288045147 cites W2928861204 @default.
- W4288045147 cites W2969268731 @default.
- W4288045147 cites W3012966247 @default.
- W4288045147 cites W3029981522 @default.
- W4288045147 cites W3094232626 @default.
- W4288045147 cites W3098913677 @default.
- W4288045147 cites W3195689905 @default.
- W4288045147 doi "https://doi.org/10.1016/j.ygeno.2022.110439" @default.
- W4288045147 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/35905834" @default.
- W4288045147 hasPublicationYear "2022" @default.
- W4288045147 type Work @default.
- W4288045147 citedByCount "8" @default.
- W4288045147 countsByYear W42880451472022 @default.
- W4288045147 countsByYear W42880451472023 @default.
- W4288045147 crossrefType "journal-article" @default.
- W4288045147 hasAuthorship W4288045147A5005315474 @default.
- W4288045147 hasAuthorship W4288045147A5007493680 @default.
- W4288045147 hasAuthorship W4288045147A5013237430 @default.
- W4288045147 hasAuthorship W4288045147A5014890138 @default.