...Scharenberg 12251 pMDLg/pRRE 3rd Packaging Contains Gag and Pol; can be used with 3rd generation transfer plasmids... Rev Trono 12260 psPAX2 2nd Packaging Contains Gag, Pol, Rev, and Tat; can be used with 2nd and 3rd generation...Trono 12263 pCMV delta R8.2 2nd Packaging Contains Gag, Pol, Rev, and Tat; can be used with 2nd and 3rd generation...Weinberg 8455 pCMV-dR8.2 dvpr 2nd Packaging Contains Gag, Pol, Rev, Tat, Vif, and Nef; can be used with 2nd ...plasmid Kasparov 22500 pNHP 2nd Packaging Contains gag, pol, tat, rev Kasparov 1732 pCPRDEnv 2nd Packaging...Packaging part of the FELIX vector system, expresses Gag-Pol and Rev Nolan 35617 pCAG-Eco N/A Envelope Ecotropic...Reiser 17531 pCD/NL-BH*DDD 2nd Packaging Contains Gag/Pol, Tat, Rev. Use with envelope plasmids from Jakob...

...MoMLV gag and pol; use with envelope plasmid Weinberg 12371 pCL-Eco Packaging Contains MoMLV gag, pol,...pol, and env Verma 35614 pBS-CMV-gagpol Packaging MoMLV gag and pol Salmon 8454 pCMV-VSV-G Envelope Envelope...

...Packaging genes (viral Gag-Pol): Gag is a structural precursor protein, and Pol is a polymerase. Envelope gene...γ-retroviruses use the same gene products for packaging (Gag, Pol, and Env), the isoforms of these proteins, as ...utilizes a packaging cell line that stably expresses Gag-Pol and/or Env, eliminating the need to deliver these...Garry Nolan at Stanford (Swift, 2001) contains Gag-Pol and either an ecotropic envelope, Phoenix-ECO, ...days. Another variant, Phoenix-gp, contains only Gag-Pol and enables additional flexibility in pseudotyping...lentiviruses and standard γ-retroviruses use the Gag, Pol, and Env genes for packaging; however, they are...Pseudotyped to alter infectivity. Packaging Gag Precursor structural protein of the retroviral ...

...MLV Gag protein. After Gag-Cas9 is incorporated, the MLV protease, supplied as part of the MLV Gag-Pol...Image from Mangeot et al., 2019.   MLV GAG-POL polyprotein Nanoblades are formed when viral structural...can be purified by centrifugation. Gag-Cas9 fusion protein The MLV Gag protein fused to a Flag-tagged SpCas9...structural proteins, such as the MLV protein Gag, multimerize and spontaneously assemble into particles at the...dCas9-VPR, if these variants are fused to the MLV Gag in place of SpCas9. gRNA A gRNA targeting a DNA sequence... by association with the the Cas9 portion of the Gag-Cas9 fusion protein. A single Nanoblade particle ...incubating Nanoblade particles with polybrene, a cationic polymer. Polybrene has long been used to increase...

...gamma-retroviruses use the same genes for packaging (that's gag, pol, and env), the isoforms of these proteins, as ... and a packaging plasmid encoding HIV proteins gag, pol, rev, and tat. 3rd generation systems require ...plasmid, and two packaging plasmids, one encoding gag and pol and a second encoding rev.  Secondly, 3rd generation...production (3 or 4). Third-generation vectors contain gag, pol, and rev. Second-generation contains those three...

...from the Didier Trono lab that expresses the HIV gag, pol, rev, and tet genes. It can be used with 2nd or...plasmid from the Trono lab that contains the gag and pol genes. It also requires pRSV-Rev and the envelope...

...wondered what goes into ‘making virus’ or what gag and pol stand for, we demystify all components of viral...particle to interact with the host. This is the role of Gag proteins: they contain the matrix, capsid, and nucleocapsid...needed for viral packaging, assembly, and display. Pol, the last component, contains the reverse transcriptase...The 5’ LTR and 3’ LTR, respectively, act as RNA polymerase II promoter and terminator regions and are slightly...replication, with E2B being the primary viral polymerase. E4/E1A activate transcription of other relevant...

...plasmid (envelope plasmid), and the polymerase (Pol) for replication and Gag structural protein on yet a third...cell line with those particles. Since the Env, Gag, and Pol genes are not encoded in the transfer plasmid... be disposed of following your lab’s biohazard policies. Administering Virus Many virus experiments involve... accident-prone researchers. Note that safety policies will vary slightly from organization to organization...with your specific organization/department/lab’s policies – basically, you’ll need to get trained in each...

...important for viral production: Gag (structural precursor protein), Pol (polymerase), Tat (viral transactivator... plasmids: one encoding Rev and one encoding Gag and Pol, increasing the number of recombination events...Naldini L, Blömer U, Gallay P, Ory D, Mulligan R, Gage FH, Verma IM, Trono D. Science. 1996 Apr 12;272(... L, Bernaudin F, Girot R, Dorazio R, Mulder GJ, Polack A, Bank A, Soulier J, Larghero J, Kabbara N, Dalle...lentivirus vector. Miyoshi H, Blömer U, Takahashi M, Gage FH, Verma IM. J Virol. 1998 Oct;72(10):8150-7. PubMed...

...constitutive RNA transport element lead to enhanced HIV-1 Gag expression in a context-dependent manner. Nucleic... both polyadenylation and termination. Out of those listed, the SV40 late polyA and rbGlob polyA are thought...any 3’ regulatory elements, such as the polyadenylation or poly(A) signal. While many studies focus on ...purpose and mechanism of polyadenylation vary across cell types, but polyadenylation generally serves to promote...consensus sequence for the addition of a poly(A) tail, the addition of a poly(A) tail on a prokaryotic transcript... termination and polyadenylation Unlike prokaryotes that have a single RNA polymerase for transcription...transcription, eukaryotes have three RNA polymerases (Polymerases I, II, and III), each responsible for transcribing...

...path to science policy and science communication A career in science communication or policy requires broad... is becoming ever more urgent for scientists to engage broadly with diverse communities, to raise awareness...and concerns of these communities. This kind of engagement and two-way dialogue is crucial if we as a society... space of science education, communication, and policy. In this post, I hope to tell you a little more...law enforcement, continues to hamper efforts to engage these communities, including those by researchers...technologies and as societal participants in the political process. My role at pgEd I have always been very...attending seminars in bioethics, public health, science policy, and science and technology studies (STS). I also...

...necessary for the production of a viral particle (Gag, Pol, Tat, Rev and Env) and the transfer plasmid encoding...because this vector requires only three HIV-1 genes (gag, pol, and rev) for production. Third generation lentiviral... you will find will recommend using a polycation, like polybrene (protocol) or DEAE dextran (protocol)...

...incorporated into HIV-1 particles independently of the Gag/Pol precursor protein. The EMBO Journal, 16(16), 5113...viral vector particles are made while missing the polymerase gene may seem like a mystery. But it’s simple... M., & Bishop, J. M. (1973). RNA-directed DNA polymerase of Rous sarcoma virus: Initiation of synthesis...

...replication. They contain two genes: gag and pol. The pol polyprotein encodes the reverse transcriptase ...reading frames (ORFs) which often terminate with a poly(A). ORF2 encodes endonuclease and reverse transcriptase...

...Packaging genes (viral Gag-Pol): Gag is a structural precursor protein, and Pol is a polymerase. Envelope gene...use the same gene products for packaging (i.e., Gag, Pol, and Env), the isoforms of these proteins, as ...utilizes a packaging cell line that stably expresses Gag-Pol and/or Env, eliminating the need to deliver these... developed by Garry Nolan at Stanford contains Gag-Pol and either an ecotropic envelope, Phoenix-ECO, ...days. Another variant, Phoenix-gp, contains only Gag-Pol and enables additional flexibility in pseudotyping...lentiviruses and standard γ-retroviruses use the Gag, Pol, and Env genes for packaging; however, they are...protein; Pseudotyped to alter infectivity. Packaging Gag in trans Precursor structural protein of the retroviral...

...Plasmid All on one plasmid: Gag, Pol, Rev, Tat Two plasmids: one encoding Gag and Pol and another encoding Rev...contains a single packaging plasmid encoding the Gag, Pol, Rev, and Tat genes. The transfer plasmid contains... plasmids: one encoding Rev and one encoding Gag and Pol. Although safer, this system can be more cumbersome... generation packaging systems: express the HIV gag, pol, rev, and tat genes all from a single packaging... . 3rd generation packaging systems: express gag and pol from one packaging plasmid and rev from another...lentiviruses and standard retroviruses use the gag, pol, and env genes for packaging; however, they are... to drive RNA pol III-directed transcription of shRNAs. cDNA expression requires RNA pol II, and thus ...

...for science policy folks is on Twitter- start an account and start engaging with the #scipol hashtag. Often... science policy, there is often discussion of “big P” and “little p” policy.  “Big P” policy is what you...about where science policy happens.  When do you do science policy? Many science policy training programs... is science policy?”  I have been asked this many times throughout my short science policy career and ...importantly, how, of a science policy career.  Who does science policy? I’ve intentionally started with... many science policy professionals that entered the field from a more traditional policy background, learning...to start. What is science policy? THE IMPOSSIBLE QUESTION. Science policy is the work you do to creatively...

... mCherry T7 Bacterial Sauka-Spengler 119971 GAG-CRErec GAG-Cre fusion; for the production of VLPs loaded...coexpression CAG Retroviral Gage 49054 CAG-GFP/cre Cre-GFP fusion CAG Retroviral Gage 49056 AAV-GFP/Cre Cre-... metallothionein MT-I region including the polyadenylation site and several introns tet-hCMV Mammalian...Cre-GFP fusion CMV AAV Gage 49111 pEMS1980 iCre with MCS for inserting promoter none AAV Simpson 49455 pSH62...Yang 107313 AAV-aCamkII-mCherry-P2A-Cre-WPRE-BGH-polyA mCherry and Cre; expressed in excitatory neurons...

...toolbox of processing factors, including polymerases (Pol μ and Pol λ), nucleases (Artemis), and structure-specific...smegmatis (Yan et al., 2020). NHEJ can also be engaged by variants of the canonical Cas9 approach. A pair...

...OPEN gAGCATCACCtcgctGGAGGTGAGc PTRF/Cavin1 OZ513 and OZ514 OPEN OPEN gACAGCCGTCtgcatctGGAGGTGCCc pitrm1...OPEN OPEN aGACTCCGTCccgtcgtGCTGAGGGCc hnf1a OZ521 and OZ522 OPEN OPEN aACCTTCCCCagagaGAGGTAGTGg VDAC2 OZ523...cACCCTCCTCgtggtGCTGGTGGCc apoeb (apolipoprotein Eb) OZ529 and OZ530 OPEN OPEN gCCCCTCAGCccagatgGGAGGAGATg dpf2 OZ531 and...OPEN OPEN tGTCTCCGGCagaggGAGGAGGAGa esrrga OZ563 and OZ564 CODA CODA tAGCTACCGCTGGCGTCTGAGGAGc HACE1 OZ565...CODA CODA gCTCATCTCCACCGAGCTGGAGAAg atp2c1 OZ567 and OZ568 CODA CODA tTACCTCCACGATCTTGCTGATGGAg dio2 OZ569...CODA OPEN cAACCTCAGCATAAAGCTGGAGGAg psen2 OZ577 and OZ578 CODA OPEN cGGCATCTCCAGAGCCCGAGGCGGACc zgc:162148...OPEN OPEN cAGCACCAGCCAGCCCAGAGTTGGGGt yap1 OZ589 and OZ590 CODA OPEN gCCCAGCTGCAGGGAGGCAGGTGAGg pdeb3 OZ591...