New and Upcoming Viral Vectors - May 2020

Since the beginning of our viral service in 2016, we’ve added many new tools to our inventory of ready-to-use viral vectors. Here are some of the AAV and lentiviral preps we have released in the last few months:

• Lentiviral preps for expressing SARS-CoV-2 proteins
• ChR2 for targeted expression
• DREADDs
• GFP dependent Cre and FLPo
• And more! 

---

New lentiviral preps

SARS-CoV-2 protein expression

The Krogan lab quickly shared plasmids expressing SARS-CoV-2 proteins at the beginning of April. Now, we've turned some of the most requested plasmids into lentiviral preps to speed up COVID-19 research even further. The following lentiviral preps are now available:

• pLVX-EF1alpha-SARS-CoV-2-N-2xStrep-IRES-Puro: Expression of the SARS-CoV-2 N protein 
• pLVX-EF1alpha-SARS-CoV-2-M-2xStrep-IRES-Puro: Expression of the SARS-CoV-2 M protein
• pLVX-EF1alpha-SARS-CoV-2-orf3a-2xStrep-IRES-Puro: Expression of the SARS-CoV-2 ORF3a

Lentiviral preps for expressing the E protein (pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro) and NSP2 (pLVX-EF1alpha-SARS-CoV-2-nsp2-2xStrep-IRES-Puro) are also in the works in and should be come available in the next few weeks.

CRISPRi pooled library

The Dolcetto pooled library is now available as a lentiviral prep. It's a human CRISPR inhibition library that inhibits over 18,000 human genes and is used for genome-wide inhibition screening.

Find the Dolcetto lentiviral prep here

Control AAV

Control AAV allow researchers to not only interpret their results, but also confirm expression in target cells. While the various AAV serotypes can be used to achieve different patterns of expression, researchers often need to determine these transduction patterns empirically for their own experiments. To this end, we offer control AAV in smaller aliquots of 20ul for serotype testing to help you pick the best AAV for your experiment. The Addgene viral service has now expanded to offer many of our control plasmids in new serotypes, as well as a new Cre dependent GFP control plasmid for expression in forebrain GABA-ergic interneurons (83895).

See our Serotype Testing AAV inventory for the entire collection!

Plasmid	Serotype	Name	Depositor
27056	AAVrg	pAAV-Ef1a-DIO EYFP	Karl Deisseroth
11447	AAV5	pAAV-Ef1a-fDIO mCherry	Karl Deisseroth
112677	AAV2	pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP	Brandon Harvey
50457	AAV2	pAAV-hSyn-DIO-EGFP	Bryan Roth
50459	AAV1	pAAV-hSyn-DIO-mCherry	Bryan Roth
50457	AAV1	pAAV-hSyn-DIO-EGFP	Bryan Roth
114472	AAVrg	pAAV-hSyn-mCherry	Karl Deisseroth
83895	AAVrg	pAAV-hDlx-Flex-GFP-Fishell_6	Gordon Fishell

Optogenetics

Optogenetics continues to supply neuroscience with ever more efficient tools for light mediated neuronal control. The advantages of optogenetics lie in the spatiotemporal control offered by the system that allows for specific manipulation of neurons. We have expanded our AAV collection to include two new ChR2 constructs that allow for targeted expression. Plasmid 75470 is now available in AAV1 for Flp-dependent expression of ChR2(H134R) bound to an mCherry reporter. We now also offer AAVrg for plasmid 83898, which targets expression of ChR2 to forebrain GAB-ergic interneurons through the mDlx enhancer element.

Plasmid	Serotype	Name	Depositor
75470	AAV1	pAAV-CAG-FLEXFRT-ChR2(H134R)-mCherry	Connie Cepko
83898	AAVrg	pAAV-mDlx-ChR2-mCherry-Fishell-3	Gordon Fishell

Chemogenetics

Chemogenetics offers noninvasive methods for manipulating cell activity, particularly in the brain, and holds promise in the development of future therapies. Our popular plasmid encoding a Cre dependent version of the DREADD hM3D is now also offered as AAV1 (along with AAV PHP.eB, AAV Retrograde, AAV2, AAV5, AAV8, and AAV9). Once activated, this DREADD acts through the Gq pathway and promotes neuronal firing.

Plasmid	Serotype	Name	Depositor
44361	AAV1	pAAV-hSyn-DIO-hM3D(Gq)-mCherry	Bryan Roth

Recombinases

Recombinase systems such as Cre-lox allow for more accurate cell and tissue targeting. While promoters and AAV serotypes can sometimes be used to target tissue types, recombinases allow you to further restrict gene expression to areas in which the recombinase and construct are coexpressed. Three new recombinases from the Cepko Lab are now available as AAV1. CRE-DOG constructs 69570 and 69571 together encode the N and C terminal segments respectively of a GFP dependent Cre. The two segments come together in the presence of GFP to form a functional Cre enzyme that induces recombination of lox sites. This system takes advantage of the extensive supply of transgenic reporter lines that supply a variety of GFP expression patterns, thus allowing you to further restrict gene activation to cells expressing both Cre and GFP. The GFP dependent FLPo, Flp-DOG (75469), is also available in AAV1.

Plasmid	Serotype	Name	Depositor
69570	AAV1	pAAV-EF1a-N-CretrcintG	Connie Cepko
69571	AAV1	pAAV-EF1a-C-CreintG	Connie Cepko
75469	AAV1	pAAV-EF1a-Flp-DOG-NW	Connie Cepko

Viral vectors coming soon!

These vectors listed below should be released at Addgene in the next 6 months, pending quality control. For an estimate on availability date or to be notified when a particular catalog item is available for order please email help@addgene.org with the vector name and serotype you’d like to be notified about.

Chemogenetics

Plasmid	Serotype	Name
44362	AAV1	pAAV-hSyn-DIO-hM4D(Gi)-mCherry
83896	AAVrg	pAAV-hDlx-GiDREADD-dTomato-Fishell-5
83897	AAVrg	pAAV-hDlx-GqDREADD-dTomato-Fishell-4

Controls

Plasmid	Serotype	Name
114471	AAV2	pAAV-Ef1a-fDIO mCherry
83900	AAVrg	pAAV-mDlx-GFP-Fishell-1

Biosensors

Plasmid	Serotype	Name
83899	AAVrg	pAAV-mDlx-GCaMP6f-Fishell-2

---

 Additional resources on the Addgene blog

• Read all of our viral vectors blog posts
• Find tips for preparing AAVs
• Download our viral vectors 101 eBook

Resources on Addgene.org

• Learn about Addgene's viral service
• Find viral protocols and videos
• Find AAVs for your research