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How to Select Promoters and Serotypes in Neurological Research?

Currently, gene therapy has been applied to the treatment of a variety of diseases, utilizing viral vectors to deliver genetic material around the organ and achieve specific expression in the target cells, thus enabling gene therapy. In neuroscience research, especially in vitro experiments, adeno-associated viral tool vectors are usually utilized to study the functional roles of genes, so how can AAV be utilized more efficiently for gene transduction in the nervous system?

Creative Biolabs presents selection strategies for AAV in neurological research. As a partner, we offer the following related services to help accelerate the progress of your program.

Our Services Descriptions
Viral Vectors Creative Biolabs has long-term devoted to the development and application of various viral vectors for neuroscience research. Based on our advanced platforms and extensive experience, we have finished a lot of challenging work.
Gene Therapies Study As an innovative company in the field of neuroscience, Creative Biolabs has successfully developed advanced gene editions and delivery technologies, which provide a powerful platform for gene therapy.
Gene-depends Neurological Disease Solutions Creative Biolabs provides our clients around the world with gene delivery, editing, modification, improvement and silencing services through advanced and comprehensive approaches, as well as design, selection and validation of your gene therapy methods.

Selection of AAV Serotypes

It has been reported that there are more than ten AAV serotypes applicable to the nervous system, such as 1, 2, 5, 6, 8, 9, PHP.B, PHP.eB, PHP.S, DJ, Retro, rh10, etc. Different serotypes have different infection efficiencies and spreading abilities in different parts of the nervous system, and the appropriate serotypes are related to the ability of the genes to achieve highly efficient and stable expression, and even related to the ultimate Therefore, it is important to select the appropriate AAV serotype by combining the labeled cells and the characteristics of each AAV serotype.

AAV Serotype Descriptions
Reversible tracer serotype - AAV2-retro AAV2-retro, which is a vector capable of effective retrograde labeling of neural loops in the mouse brain with retrograde tracing ability and efficiency comparable to that of conventional retrograde tracers.
Serotypes that can cis-span single-level synapses - AAV1 AAV1 is able to propagate cis-synaptically across synapses under high titer conditions and, in combination with a Cre-dependent reporter system, efficiently and specifically drives Cre-dependent expression of exogenous genes in postsynaptic neurons for labeling postsynaptic neurons.
Serotypes that can cross the blood-brain barrier - AAV, AAVPHP.B, AAVPHP.eB, AAVPHP.S, AAV.CPP.16 AAV9 & AAVPHP.B & AAVPHP.eB & AAVPHP.S can effectively mediate the crossing of genes from the periphery to the central nervous system, followed by delivery of genetic material to the brain or spinal cord.
Serotype that efficiently targets neuronal cells - AAV.CAP-B10 AAV. CAP-B10 exhibits specificity for the central nervous system, with high targeting of neuronal cells and low targeting of the liver.
Serotype for efficient targeting of astrocytes - rAAV11 rAAV11 can efficiently target projection neurons retrogradely and does not exhibit cis-trans-synaptic characteristics. rAAV11 has excellent retrograde labeling efficiency. rAAV11 also has excellent retrograde labeling efficiency and can trace projection neurons that are difficult to transduce with rAAV2.
Serotypes that efficiently target microglia - AAV-cMG & AAV-MG AAV-MG1.1 and AAV-MG1.2, which can efficiently invade microglia in vivo without inducing microglia immune activation. AAV-cMG is suitable for in vitro invasion of cultured microglia and AAV-MG is suitable for in vivo invasion of endogenous microglia.
Serotypes efficiently targeting central vascular endothelial cells - AAV-BI30 AAV-BI30 is efficiently and specifically expressed in different types of central vascular endothelial cells, which is suitable for research related to neurovascular biology and contributes to the development of therapeutic approaches for endothelial dysfunction diseases.

A number of AAV products are important tools commonly used in research. You can browse the table below to see a list of our recommended products.

Cat. No Product Name Types
NTC2101070CR AAV2 Full Capsids, Reference Standards Reference Materials
NRZP-0722-ZP719 rAAV-CAG-DIO-G-Flamp1 Fluorescent cAMP Indicators.
NTA-2012AD-P508 pAAV-hSyn-DIO-XCaMP-R-WPRE Calcium Indicator
NRZP-0622-ZP616 AAV-EF1a-mCherry-flex-dtA Cell Ablation
NTA-2012-ZP303 AAV2/2Retro-CAG-DIO-EGFP-2A-TetTox-pA Retrograde Tracing
NTA-2012-ZP268 AAV2/9-hEF1a-DIO-mCherry-P2A-TetTox-WPRE-pA Synaptic Inactivation
NTA-2012-ZP78 AAV2/9-hEF1a-fDIO-eNpHR 3.0-mCherry-WPRE-pA Chemogenetic Inhibition
NRZP-0622-ZP721 AAV2/9-mDLX-iCRE-tdTomato-WPRE-pA Cre Recombinase
NRZP-0722-ZP719 rAAV-CAG-DIO-G-Flamp1 Neurotransmitter Sensor
NTA-2012-ZP149 AAV2/9-hSyn-Flpo-EGFP-WPRE-pA Neural Tracing
NTA-2011-ZP20 VSV-eGFP [Vesicular Stomatitis Virus (VSV)] Neural Tracing

Selection of Promoters

In addition to AAV vectors and serotypes, optimization of promoters and regulatory elements is also necessary. Currently, broad-spectrum type promoters used in the study, such as

  • Cytomegalovirus promoter CMV and CMV enhancer fused chicken ß-actin promoter CAG can achieve high expression of target genes.
  • The tetracycline promoter TRE acts as an inducible promoter and can initiate gene expression under specific conditions.

However, the composition of the nervous system is extremely complex, and sophisticated studies of the brain are critical, and it is particularly important to select a promoter with relatively higher specificity in order to achieve specific infection of a certain brain region or even a certain class of cells.

The following table shows some common neural tissue-specific promoters.

Promoter Name Promoter Size Promoter Source Promoter Applications
hSyn 471 bp Human Neuron-specific promoters
CamKIla 1.2 kb Mice Neuron-specific promoters of excitatory neurons in the neocortex and hippocampus of the brain
c-fos 1.7 kb Mice Excitatory neuron promoters
Mecp2 230 bp Mice Short neuron-specific promoters
NSE 1.3 kb Mice Neuron-specific promoters
Somatostat (SST) 1.2 kb Human gamma aminobutyric acidergic inhibitory neuron SST subtype-specific promoter
TH 2.5 kb Rat Dopaminergic neuron-specific promoter
GFAP 2.0 kb Human Astrocyte-specific promoter
GFAP104 845 bp Human astrocyte-specific promoter
GfaABC1D(truncated GFAP) 681 bp Human Astrocyte-specific promoters
ALDHIL1 1.3 kb Human Astrocyte-specific promoters in the thalamus
MBP 1.3 kb Human Oligodendrocyte-specific promoter
mDlx 530 bp Mice GABAergic interneuron-specific promoter
GAD65 2.5 kb Mice GABAergic neuron/interneuron specific promoter
ePet-1 1.9 kb Mice Serotonin (5-light tryptamine)-ergic neuron-specific promoter
hSyn 471 bp Human Neuron-specific promoters

Therefore, to drive more efficient gene transduction in neurological studies, all of these factors can be taken into account to optimize the choice.

For Research Use Only. Not For Clinical Use.
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