New and Upcoming Viral Vectors - September 2019

By Leila Haery

New Viral Vectors AddgeneSince 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 we have released in the last few months. You can also browse our entire AAV inventory.

Our new AAVs include:

  • EGFP-expressing AAV for serotype testing
  • Calcium sensors and GABA sensors
  • Optogenetics
  • Recombinases
  • And more!


Control AAV

For many AAV experiments, we recommend validating the serotype in your target cells before conducting the experiment. Likewise, a lot of scientists have requested samples of AAV for testing purposes. To address this, we recently released our serotype testing AAV, which are small (20 ul) samples of EGFP-expressing AAV packaged in various serotypes. This collection is now complete! pAAV-CAG-GFP (plasmid 37825) and pAAV-hSyn-EGFP (plasmid 50465) are now available as 20 ul aliquots in AAV1, AAV2, AAV5, AAV8, AAV9, and AAV Retrograde. You can also request the specific serotypes you need instead of receiving a kit with all the serotypes, since these 20 ul samples are distributed individually.

See our Serotype Testing AAV inventory for the entire collection!

We are expanding our entire control AAV inventory to give scientists access to a wide array of promoters and fluorophores for validating and testing purposes. We will continue to add more controls, and have recently released these controls:

  • pAAV-hSyn-DIO-EGFP (50457-AAV5): The Synapsin promoter directs broad, neuronal expression.
  • AAV pCAG-FLEX-EGFP-WPRE (51502-AAV5): The CAG promoter directs ubiquitous expression.
  • pAAV-mDlx-NLS-mRuby2 (99130-AAVrg): The Dlx element directs expression in forebrain GABAergic interneurons.
  • pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP (112677-AAV5 and 112677-AAVrg): The EF1a promoter directs strong, ubiquitous expression. This is a Cre-dependent switch that expresses nuclear mCherry in the absence of Cre, and expresses nuclear EGFP in the presence of Cre.

Biosensor AAV (calcium sensors and GABA sensors)

Calcium sensors

The latest calcium sensors from Janelia (jGCaMP7) continue to be useful for monitoring neuronal activity in the form of calcium dynamics. (For more details on this technology, see our blog post Overview of AAV Encoded Calcium Sensors). We are expanding the inventory of these vectors to more serotypes. We currently package four variants of these biosensors, which are optimized for specific parameters:

  • jGCaMP7s is the most sensitive responder to action potentials. For a single action potential, GCaMP7s is approximately twice as bright, relative to baseline fluorescence, as the GCaMP7f variant, and is approximately 5-fold brighter than GCaMP6s.
  • jGCaMP7f is the fastest responding variant.
  • jGCaMP7b exhibits the brightest resting fluorescence and can be used for imaging small neuronal processes (dendrites and axons).
  • jGCaMP7c exhibits high contrast between peak fluorescence and resting fluorescence (achieved through lowering sensor resting fluorescence, while maintaining high peak fluorescence) and is useful for imaging signaling activity in large populations of densely-labeled neurons because background fluorescence from inactive neurons is reduced. This variant has an approximately 12-fold increase in brightness after ~100 action potentials. This is about twice as bright as GCaMP7f and GCaMP7s.

See all our biosensors inventory, and check out our latest vectors below:

GABA sensors

Sensing levels of g-Amino butyric acid (GABA) - the primary inhibitory neurotransmitter - is a powerful method for understanding how this molecule impacts neuronal circuits. We recently packaged a constitutive GABA sensor (see below), in addition to our inventory of Cre-dependent GABA sensors. See all our GABA sensor AAV inventory here.

Optogenetics

Optogenetics continues to be a useful method for manipulating specific neurons. We recently packaged a soma-targeted opsin from Christopher Harvey’s lab. According to their publication (Chettih & Harvey, 2019), “this localization should improve the specificity of influence measurements by reducing photostimulation of the axons and dendrites of non-targeted neurons near the target site.”

Click here to find viral vectors at Addgene

Recombinase AAV

Targeting and manipulating specific, genetically-defined neuronal populations with intersectional approaches is a growing field that we aim to support by growing our inventory of recombinases and recombinase-dependent vectors. We recently released several recombinases in various serotypes, and will continue to grow this collection. Of note, we are excited to release a Flp-dependent Cre AAV in AAV9 and AAVrg. See our new additions here:

Cre vectors

Dre vector

Flp vectors

Viral vectors coming soon!

These vectors should be packaged and available as AAV in the next few months. Email us or check our website to access them!

Controls

  • pAAV-hSyn-hChR2(H134R)-EYFP (26973-AAVrg)
  • pAAV-hSyn-hChR2(H134R)-mCherry (26976-AAV5)
  • pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP (112677-AAV5)

Calcium Sensors

  • pGP-AAV-syn-jGCaMP7b-WPRE (104489-AAVrg)
  • pGP-AAV-syn-FLEX-jGCaMP7b-WPRE (104493-AAVrg)
  • pGP-AAV-CAG-FLEX-jGCaMP7s-WPRE (104495-AAVrg)
  • pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE (104496-AAVrg)

Optogenetics

  • pAAV-hSyn Con/Fon hChR2(H134R)-EYFP (55645-AAV5)

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 Additional resources on the Addgene blog

Resources on Addgene.org

Topics: Viral Vectors, Addgene’s Viral Service

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