Matches in SemOpenAlex for { <https://semopenalex.org/work/W4386293868> ?p ?o ?g. }
- W4386293868 abstract "Summary/Abstract Non-viral DNA donor template has been widely used for targeted genomic integration by homologous recombination (HR). This process has become more efficient with RNA guided endonuclease editor system such as CRISPR/Cas9. Circular single stranded DNA (cssDNA) has been harnessed previously as a g enome engineering c atalyst (GATALYST) for efficient and safe targeted gene knock-in. However, the engineering efficiency is bottlenecked by the nucleoplasm trafficking and genomic tethering of cssDNA donor, especially for extra-large transgene integration. Here we developed enGager, en hanced G ATALYST a ssociated g enome e ditor system by fusion of nucleus localization signal (NLS) peptide tagged Cas9 with various single stranded DNA binding protein modules through a GFP reporter Knock-in screening. The enGager system assembles an integrative genome integration machinery by forming tripartite complex for engineered nuclease editors, sgRNA and ssDNA donors, thereby facilitate the nucleus trafficking of DNA donors and increase their active local concentration at the targeted genomic site. When applied for genome integration with cssDNA donor templates to diverse genomic loci in various cell types, these enGagers outperform unfused editors. The enhancement of integration efficiency ranges from 1.5- to more than 6-fold, with the effect being more prominent for > 4Kb transgene knock-in in primary cells. We further demonstrated that enGager mediated enhancement for genome integration is ssDNA, but less dsDNA dependent. Using one of the mini-enGagers, we demonstrated large chimeric antigen receptor (CAR) transgene integration in primary T cells with exceptional efficiency and anti-tumor function. These tripartite e ditors with s sDNA o ptimized g enome en gineering system (TESOGENASE TM ) add a set of novel endonuclease editors into the gene-editing toolbox for potential cell and gene therapeutic development based on ssDNA mediated non-viral genome engineering. Highlight A reporter Knock-in screening establishes enGager system to identify TESOGENASE editor to improving ssDNA mediated genome integration Mini-TESOGENASEs developed by fusing Cas9 nuclease with novel ssDNA binding motifs mRNA mini-TESOGENASEs enhance targeted genome integration via various non-viral delivery approaches Efficient functional CAR-T cell engineering by mini-TESOGENASE" @default.
- W4386293868 created "2023-08-31" @default.
- W4386293868 creator A5011231000 @default.
- W4386293868 creator A5011700288 @default.
- W4386293868 creator A5012842307 @default.
- W4386293868 creator A5016123511 @default.
- W4386293868 creator A5026703614 @default.
- W4386293868 creator A5027765985 @default.
- W4386293868 creator A5028421783 @default.
- W4386293868 creator A5037411637 @default.
- W4386293868 creator A5089734920 @default.
- W4386293868 date "2023-08-29" @default.
- W4386293868 modified "2023-09-28" @default.
- W4386293868 title "TESOGENASE, An Engineered Nuclease Editor for Enhanced Targeted Genome Integration" @default.
- W4386293868 cites W1091016337 @default.
- W4386293868 cites W1513848044 @default.
- W4386293868 cites W2003171404 @default.
- W4386293868 cites W2006654626 @default.
- W4386293868 cites W2012774405 @default.
- W4386293868 cites W2036099589 @default.
- W4386293868 cites W2041929141 @default.
- W4386293868 cites W2050480410 @default.
- W4386293868 cites W2064815984 @default.
- W4386293868 cites W2066783444 @default.
- W4386293868 cites W2077904712 @default.
- W4386293868 cites W2080924650 @default.
- W4386293868 cites W2096261947 @default.
- W4386293868 cites W2098110766 @default.
- W4386293868 cites W2098865666 @default.
- W4386293868 cites W2101749257 @default.
- W4386293868 cites W2116516380 @default.
- W4386293868 cites W2152208990 @default.
- W4386293868 cites W2160051211 @default.
- W4386293868 cites W2336828812 @default.
- W4386293868 cites W2344793489 @default.
- W4386293868 cites W2521981506 @default.
- W4386293868 cites W2547418311 @default.
- W4386293868 cites W2548694234 @default.
- W4386293868 cites W2589417329 @default.
- W4386293868 cites W2590493137 @default.
- W4386293868 cites W2593664217 @default.
- W4386293868 cites W2608534885 @default.
- W4386293868 cites W2755829020 @default.
- W4386293868 cites W2767634557 @default.
- W4386293868 cites W2768823259 @default.
- W4386293868 cites W2769263426 @default.
- W4386293868 cites W2818540548 @default.
- W4386293868 cites W2935121891 @default.
- W4386293868 cites W2942076130 @default.
- W4386293868 cites W2945532708 @default.
- W4386293868 cites W2951185264 @default.
- W4386293868 cites W2955483819 @default.
- W4386293868 cites W2966261467 @default.
- W4386293868 cites W2992036306 @default.
- W4386293868 cites W3000874218 @default.
- W4386293868 cites W3001754284 @default.
- W4386293868 cites W3081421945 @default.
- W4386293868 cites W3102945151 @default.
- W4386293868 cites W3106947848 @default.
- W4386293868 cites W3109058715 @default.
- W4386293868 cites W3119901427 @default.
- W4386293868 cites W3120704519 @default.
- W4386293868 cites W3121885986 @default.
- W4386293868 cites W3133746937 @default.
- W4386293868 cites W3134798812 @default.
- W4386293868 cites W3161425918 @default.
- W4386293868 cites W3168477404 @default.
- W4386293868 cites W3170899528 @default.
- W4386293868 cites W3177445623 @default.
- W4386293868 cites W3188940990 @default.
- W4386293868 cites W3191779858 @default.
- W4386293868 cites W3206678867 @default.
- W4386293868 cites W3206987978 @default.
- W4386293868 cites W4210413600 @default.
- W4386293868 cites W4224255508 @default.
- W4386293868 cites W4226211972 @default.
- W4386293868 cites W4232090872 @default.
- W4386293868 cites W4289939894 @default.
- W4386293868 cites W4293237445 @default.
- W4386293868 cites W4293792836 @default.
- W4386293868 cites W4294911599 @default.
- W4386293868 cites W4307996358 @default.
- W4386293868 doi "https://doi.org/10.1101/2023.08.28.553855" @default.
- W4386293868 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/37693500" @default.
- W4386293868 hasPublicationYear "2023" @default.
- W4386293868 type Work @default.
- W4386293868 citedByCount "0" @default.
- W4386293868 crossrefType "posted-content" @default.
- W4386293868 hasAuthorship W4386293868A5011231000 @default.
- W4386293868 hasAuthorship W4386293868A5011700288 @default.
- W4386293868 hasAuthorship W4386293868A5012842307 @default.
- W4386293868 hasAuthorship W4386293868A5016123511 @default.
- W4386293868 hasAuthorship W4386293868A5026703614 @default.
- W4386293868 hasAuthorship W4386293868A5027765985 @default.
- W4386293868 hasAuthorship W4386293868A5028421783 @default.
- W4386293868 hasAuthorship W4386293868A5037411637 @default.
- W4386293868 hasAuthorship W4386293868A5089734920 @default.
- W4386293868 hasBestOaLocation W43862938681 @default.
- W4386293868 hasConcept C102230213 @default.
- W4386293868 hasConcept C102744134 @default.