.png){kind=icon}
Return
Split selectable markers
.png){kind=icon}
OA
AI
DOI：10.1038/s41467-019-12891-2.png)
.png){kind=icon}
.png){kind=icon}
Share.png){kind=icon}
Save
Abstract
En 中文
Selectable markers are widely used in transgenesis and genome editing for selecting engineered cells with a desired genotype but the variety of markers is limited. Here we present split selectable markers that each allow for selection of multiple unlinked transgenes in the context of lentivirus-mediated transgenesis as well as CRISPR-Cas-mediated knock-ins. Split marker gene segments fused to protein splicing elements called inteins can be separately co-segregated with different transgenic vectors, and rejoin via protein trans-splicing to reconstitute a full-length marker protein in host cells receiving all intended vectors. Using a lentiviral system, we create and validate 2-split Hygromycin, Puromycin, Neomycin and Blasticidin resistance genes as well as mScarlet fluorescent proteins. By combining split points, we create 3- and 6-split Hygromycin resistance genes, demonstrating that higher-degree split markers can be generated by a chaining design. We adapt the split marker system for selecting biallelically engineered cells after CRISPR gene editing. Future engineering of split markers may allow selection of a higher number of genetic modifications in target cells.
Keywords:
DNAE INTEIN
Journal
IF:
15.7
Papers:
9.6W
Citations:
93.2W
Organization
Cited Papers
Highly efficient protein trans-splicing by a naturally split DnaE intein from Nostoc punctiforme.png)
FEBS LETTERS
OAAI
FEBS LETTERSIF3
OAAIShareSaveThe naturally split Npu DnaE intein exhibits an extraordinarily high rate in the protein trans-splicing reactionFEBS LETTERS
FEBS LETTERSIF3
PREAI
ShareSaveThe human-induced pluripotent stem cell initiative-data resources for cellular geneticsNUCLEIC ACIDS RESEARCHOAAI
NUCLEIC ACIDS RESEARCHIF13.1
OAAIShareSaveShareSaveShareSaveFaster Protein Splicing with the Nostoc punctiforme DnaE Intein Using Non-native Extein ResiduesOAAI
OAAIShareSaveUnprecedented Rates and Efficiencies Revealed for New Natural Split Inteins from Metagenomic SourcesOAAI
OAAIShareSave
View more