Programmable protein delivery with a bacterial contractile injection system.

Kreitz J, Friedrich MJ, Guru A, Lash B, Saito M, Macrae RK, Zhang F
Nature. 2023 Apr;616(7956):357-364. doi: 10.1038/s41586-023-05870-7. Epub 2023 Mar 29. PubMed Article

Plasmids from Article

ID	Plasmid	Purpose
198271	pAWP78-PVCpnf1-16	Unmodified pPVC; contains WT PVC structural/accessory genes
198272	pBR322-PVCpnf17-22	Unmodified pPayload; contains WT PVC payloads/regulatory genes
198273	pBR322-PVCpnf18-21	Contains only regulatory genes; for generating empty PVCs
198274	pBR322-Pdp1_NTD-Cre	For loading Cre into PVCs; also contains PVC regulatory genes
198275	pBR322-Pdp1_NTD-Cas9	For loading SpCas9 into PVCs; also contains PVC regulatory genes
198276	pBR322-Pdp1_NTD-ZFD_L	For loading L half of zinc finger deaminase targeting human TRAC locus (ZFD_TRAC-NC-Left-G1397-N_DddA); also contains PVC regulatory genes
198277	pBR322-Pdp1_NTD-ZFD_R	For loading R half of zinc finger deaminase targeting human TRAC locus (ZFD_TRAC-NC-Right-G1397-C_DddA); also contains PVC regulatory genes
198278	pIEx4-Dio-GFP	Cre reporter for insect cells; double-floxxed-EGFP
198279	pcDNA3-Dio-GFP	Cre reporter for mammalian cells; double-floxxed-EGFP
198280	pIEx4-Cre	Expresses Cre in insect cells
198281	pcDNA3-Cre	Expresses Cre in mammalian cells
198282	py100-sgRNA_hVEGFA	Expression of SpCas9 guide RNA specific for human VEGFA locus
198283	pAWP78-PVCpnfΔpvc13	Produces PVCs lacking tail fibers
198284	pAWP78-PVCpnfΔpvc15	Produces PVCs lacking payloads by elimination of the payload loader (pvc15)
198285	pAWP78-PVCpnf_FLAG-pvc2	Contains an external epitope tag on the sheath protein; for immunofluorescence-based binding assays
198286	pAWP78-PVCpnf_pvc13-truncated	Contains a tail fiber with a truncated binding domain (lacking aa403-476 in Pvc13); produces nontargeting PVCs
198287	pAWP78-PVCpnf_pvc13-E01DARPin	Produces PVCs retargeted with anti-EGFR DARPin E01
198288	pAWP78-PVCpnf_pvc13-A4DARPin	Produces PVCs retargeted with anti-lysozyme DARPin A4
198289	pAWP78-PVCpnf_pvc13-Ad5Knob	Produces PVCs retargeted with knob domain from hAd5 virus
198290	pAWP78-PVCpnf_pvc13-Ad5DL491-492	Produces PVCs retargeted with non-binding mutant of Ad5 knob
198291	pAWP78-PVCpnf_pvc13-Ad5RGDPK7	Produces PVCs retargeted with expanded tropism hAd5 knob (RGD and PK7 insertions)
198292	pAWP78-PVCpnf_pvc13-antiMouseMHCIINb	Produces PVCs retargeted with VHH7 anti-mouse MHC class II nanobody
198293	pAWP78-PVCpnf_pvc13-antiHistoneNb	Produces PVCs retargeted with A9 anti-histone nanobody
198294	pAWP78-PVCpnf_pvc13-CD4DARPin	Produces PVCs retargeted with anti-CD4 DARPin D27.2
198295	pAWP78-PVCpnf_pvc13-CD11bDARPin	Produces PVCs retargeted with anti-CD11b DARPin F7
198296	pAWP78-PVCpnf_pvc13-HA	Produces PVCs retargeted with HA tag
198297	pAWP78-PVCpnf_pvc13-FLAG	Produces PVCs retargeted with FLAG tag
198298	pAWP78-PVCpnf_pvc13-EE	Produces PVCs retargeted with EE tag
198299	pAWP78-PVCpnf_pvc13-MoonTag	Produces PVCs retargeted with MoonTag
198300	pAWP78-PVCpnf_pvc13-SunTag	Produces PVCs retargeted with SunTag
198301	pAWP78-PVCpnf_pvc13-ALFA	Produces PVCs retargeted with ALFA tag
198302	pHCMV-SS-antiHAscFv-TM	Displays anti-HA scFv from the surface of mammalian cells
198303	pHCMV-SS-antiFLAGscFv-TM	Displays anti-FLAG scFv from the surface of mammalian cells
198304	pHCMV-SS-antiEEscFv-TM	Displays anti-EE scFv from the surface of mammalian cells
198305	pHCMV-SS-antiMoonTagNb-TM	Displays anti-MoonTag nanobody from the surface of mammalian cells
198306	pHCMV-SS-antiSunTag-TM	Displays anti-SunTag scFv from the surface of mammalian cells
198307	pHCMV-SS-antiALFAtagNb-TM	Displays anti-ALFA nanobody from the surface of mammalian cells
198308	pAWP78-PVCpnf_pvc12-HiBiT	Produces PVCs tagged with HiBiT on the baseplate; used as assembly/loading control for payload loading assays
198309	pAWP78-PVCpnfΔpvc15_pvc12-HiBiT	Negative control for payload loading assay; produces PVCs deficient for payload loader (pvc15)
198310	pBR322-Pdp1-HiBiT	Loads HiBiT-tagged Pdp1 into PVCs; for payload loading assays; also contains Pnf
198311	pBR322-Pdp1_aa1-289-HiBiT	Loads a truncated payload; for identification of Pdp1 packaging domain
198312	pBR322-Pdp1_aa1-190-HiBiT	Loads a truncated payload; for identification of Pdp1 packaging domain
198313	pBR322-Pdp1_aa1-133-HiBiT	Loads a truncated payload; for identification of Pdp1 packaging domain
198314	pBR322-Pdp1_aa1-76-HiBiT	Loads a truncated payload; for identification of Pdp1 packaging domain
198315	pBR322-Pdp1_aa1-38-HiBiT	Loads a truncated payload; for identification of Pdp1 packaging domain
198316	pBR322-Pdp1_aa39-331-HiBiT	Loads a truncated payload; for identification of Pdp1 packaging domain
198317	pBR322-Pdp1_NTD-GFP-HiBiT	Loads HiBiT-tagged GFP into PVCs using Pdp1 packaging domain; also contains Pnf
198318	pBR322-Pdp1_NTD-Cre-HiBiT	Loads HiBiT-tagged Cre into PVCs using Pdp1 packaging domain; also contains Pnf
198319	pBR322-Pdp1_NTD-ZFN_L-HiBiT	Loads HiBiT-tagged L fragment of a zinc finger nuclease specific for human AAVS1 locus using Pdp1 packaging domain
198320	TwinStrep-SUMO-Pdp1	For affinity purification of Pdp1 payload
198321	TwinStrep-SUMO-Pnf	For affinity purification of Pnf payload
198448	pAWP78-PVCpnf_Δpvc10	Produces PVCs containing truncated spike tip protein (Pvc10).
198449	pAWP78-PVCpnf_pvc13-Ad5RGDPK7_Δpvc10	Produces PVCs retargeted with expanded tropism hAd5 knob (RGD and PK7 insertions); also contains truncated spike tip protein (Pvc10).
198450	pAWP78-PVCpnf_pvc13-antiMouseMHCIINb_Δpvc10	Produces PVCs retargeted with VHH7 anti-mouse MHC class II nanobody; also contains truncated spike tip protein (Pvc10).