Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Filters applied. Clear all
. 2014 Aug;11(8):783-784.
doi: 10.1038/nmeth.3047.

Improved Vectors and Genome-Wide Libraries for CRISPR Screening

Affiliations
Free PMC article

Improved Vectors and Genome-Wide Libraries for CRISPR Screening

Neville E Sanjana et al. Nat Methods. .
Free PMC article

Figures

Figure 1
Figure 1
New lentiviral CRISPR designs produce viruses with higher functional titer. (a) Lentiviral expression vector for Streptococcus pyogenes Cas9 and sgRNA in the improved one vector system (lentiCRISPR v2) and the two vector system (lentiCas9-Blast, lentiGuide-Puro). Psi packaging signal (psi+), rev response element (RRE), central polypurine tract (cPPT), elongation factor-1α short promoter (EFS), FLAG octapeptide tag (FLAG), 2A self-cleaving peptide (P2A), puromycin selection marker (puro), posttranscriptional regulatory element (WPRE), blasticidin selection marker (blast), and elongation factor-1α promoter (EF1a). (b) Relative functional viral titer of viruses made from lentiCRISPR v1, lentiCRISPR v2, and lentiGuide-Puro vector with a EGFP-targeting sgRNA (n = 3 independently-transfected virus batches with 3 replicate transductions per construct). HEK293FT cells were transduced with serial dilutions of virus and, after 24 hours, selected using puromycin (1 ug/ml). Puromycin-resistant cells were measured after 4 days from the start of selection using the CellTiter-Glo (Promega) luciferase assay. Relative titers were calculated using viral volumes that yielded less than 20% puromycin-resistant cells in order to minimize the number of cells with multiple infections. Numbers above each bar indicate the size of the packaged virus for each construct.

Similar articles

See all similar articles

Cited by 1,200 articles

See all "Cited by" articles

LinkOut - more resources

Feedback