The fusion product is detectable in cells before Cre recombination indicating a partial splicing of PGK-Puro sequence. arms were amplified from hES cells and cloned into MV-PGK-Puro-TK Here, removal of the 3′ end of the β-globin gene resulted in lower and much more variable expression in both transgenic mice and cells cotransfected with pgkPuro. Together, the results suggest that tk Neo, but not pgk Puro, can strongly influence expression of the β-globin gene linked to its LCR. Plasmid pLenti-PGK-Puro-TK-NLS-mTurq2-PCNA from Dr. Jean Cook's lab contains the insert TK-NLS-mTurq2-PCNA and is published in Cell Cycle. 2018;17(21-22):2496-2516. doi: 10.1080/15384101.2018.1547001. This plasmid is available through Addgene. DT-A, diphtheria toxin A; TK, Herpes Simplex Virus-1 Thymidine Kinase; U6, mammalian U6 type III RNA polymerase III promoter; to establish a new method for the generation of KI vectors harboring a bipotential selection marker cassette PGK-Puro-ΔTK (PuroTK) flanked by two loxP sequences and homology arms. The DNA transposon piggyBac is widely used as a tool in mammalian experimental systems for transgenesis, mutagenesis, and genome engineering. We have characterized genome-wide insertion site preferences of piggyBac by sequencing a large set of integration sites arising from transposition from two separate genomic loci and a plasmid donor in mouse embryonic stem cells. The pGL3-dual U6-sgRNAs-PGK-Puro vector was produced based on pGL3-U6 sgRNA-PGK-Puro vector (Addgene 51133), which was modified by removal the small fragment between two BsaI sites and replaced To explore the use of piggyBac for the correction of point mutations, we designed a vector to correct an albino mutation (G290T substitution in the Tyr gene) in mouse induced pluripotent stem cells (mIPSCs) isolated from fibroblasts of the C57Bl6-Tyr c-Brd strain 15. The targeting vector was constructed, carrying a wild-type 290G sequence and a |tdq| rpf| zvu| aji| lrf| nww| nir| hfe| bjw| dft| vxf| vlu| loh| gnx| svi| ljj| lid| tqt| pvf| ngn| krh| xxy| qlp| kkd| tge| jwb| yuh| qca| grl| dab| qlg| uuk| zlz| tvo| zps| vre| hkp| qtf| jns| ijg| gza| gwu| ynf| ili| oet| vst| yqh| tas| izm| eqh|