...using ssDNA oligo as donor template, it's usually more efficient than plasmid donor. Typical design is ...the HR template, you usually would need to make mutations of the HR template to avoid this donor plasmid... HR template vector design guidelines? A few notes below are considerations for designing HR donor. Generally...my HR template an issue? Yes. If you have intact protospacer+PAM sequence within the HR template, it will...experiment, consider co-transfecting with GFP. This allows you to sort for GFP-positive cells and to enrich for...then select the best guide to co-transfect in HR donor. Alternatively, a new system described recently ...insertions or deletions), we typically use plasmid donor, with two homology arms on each side flanking your...

... the genomic site and the donor vector. The PITCh donor plasmid with an EGFP-2A-PuroR cassette, flanked... and donor vectors as well as pX330S-2-PITCh, expressing a generic gRNA targeting the PITCh donor plasmid...following articles provide PCR templates for amplification of the tag (eg GFP, Flag, YFP, etc) and selection...Plasmids Protein Species Tag Donor Plasmid gRNA plasmid gRNA plasmid FLAG pFETCh_Donor CEBPB Human FLAG pFETCh_CEBPB...FBL-specific gRNA pCRIS-PITChv2-FBL - PITCh donor vector for EGFP-2A-PuroR knock-in into human FBL locus Jorgensen...created using CRISPR and the donor plasmids containing homology arms and EGFP are available at Addgene. ...101 post for more information about HDR/repair template design. How to use CRISPR to tag your gene of ...

...genomic edits by copying from a DNA donor template . The donor template contains the desired sequence and...linear dsDNA templates work better for large edits or insertions. When designing the donor template, incorporating...out integration of donor DNA. Both types of CASTs have been found to achieve donor DNA integration for...pathway. HDR relies on copying DNA from a matching template to accurately repair or fill in the missing sequence...the break ends without the need for a homologous template. Both NHEJ and HDR have their uses, but we’ll ...cell along with the gRNA(s) and Cas enzyme. The donor DNA contains the desired edit as well as additional...homology arms. Depending on the application, the donor DNA may be a single-stranded oligonucleotide, double-stranded...

...activator with 2A GFP 61423 : Expresses the MS2-P65-HSF1 activator helper complex with 2A GFP 61424 : sgRNA...(SapI)_hSyn-GFP-KASH-bGH (SpGuide acceptor). This is a AAV plasmid for sgRNA cloning. GFP-KASH fusion ...contains a Kras G12D homology directed repair donor template. #60225 - AAV:ITR-U6-sgRNA(LacZ)-pEFS-Rluc-...PX552; U6 promoter-driven; for sgRNA cloning; has GFP-KASH for FACS sorting SaCas9 + single guide RNA: ...luciferase-2A-Cre recombinase, and Kras G12D HDR template 60225 : control for AAV-KPL; sgRNA targeted to... recombinase-2A-EGFP-KASH 60231 : sgRNA cloning backbone with Cre recombinase-2A-EGFP-KASH Detailed backbone...recombinase-2A-EGFP-KASH and an sgRNA. #60231 - AAV:ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR...

... pCAG-SpCas9-GFP-U6-gRNA 79144 Mammalian hU6 yes, cut S. pyogenes EGFP Zou pCAG-eCas9-GFP-U6-gRNA 79145...Cerulean Church LRG (Lenti_sgRNA_EFS_GFP) 65656 Mammalian/Lentiviral LIC none S. pyogenes GFP Vakoc U6>sgRNA...cut Lb Cpf1 Neo Welker AIO-GFP 74119 Mammalian U6x2 yes, nick S. pyogenes EGFP Jackson AIO-mCherry 74120...S. pyogenes Kamoun pUC119-gRNA 52255 Plant PCR template none S. pyogenes Sheen pRGEB31 51295 Plant BsaI...pyogenes Zhang pL-CRISPR.EFS.GFP 57818 Mammalian/Lentiviral BsmBI yes, cut S. pyogenes EGFP Ebert pLKO.1-puro...pyogenes Zhang pLKO5.sgRNA.EFS.GFP 57822 Mammalian/Lentiviral BsmBI none S. pyogenes EGFP Ebert pSECC 60820...tagRFP657 Ebert pL-CRISPR.SFFV.GFP 57827 Mammalian/Lentiviral BsmBI yes, cut S. pyogenes EGFP Ebert pLKO5.sgRNA.EFS.tRFP...