RediMODEL transgenics in C. elegans

SLC6A3 - dopamine transporter

GENE DESCRIPTION:

  • Function: SLC6a3 is an integral membrane protein responsible for transporting the dopamine neurotransmitter from synaptic space back into presynaptic neurons.
  • Disease: Genetic defects in SLC6A3 have been found to be highly associated with Infantile Parkinsonism-Dystonia.
 

Products:

RediMODEL Kits -  a series of humanized animal models specifically engineered for easy measurement of phenotype defects caused by clinically-relevant disease mutations.   

rediMODEL-KO

SLC6A3 RediMODEL KO: Generated in the C. elegans homolog dat-1 with deletion of coding sequence to create a knock-out (KO) strain. Loss of function from dat-1 deletion leads to build up of dopamine neurotransmitter at the synapse. Major phenotype readout is the observation of paralysis upon exposure to liquid with the Swimming Worm Induced Paralysis (SWIP) assay. An all-coding deletion mutant is currently available for preorder as gene knock-out.

 

SLC6A3 RediMODEL KI: Generated in the C. elegans homolog dat-1 with human gene insertion to create a knock-in (KI) strain. Entire coding of dat-1 is replaced with wild-type SLC6A3 coding sequence. A SLC6A3 gene replacement is available for preorder as a humanized wt line.

 

SLC6A3 RediMODEL CV: Generated in the C. elegans homolog dat-1 with human gene insertion containing a clinical variant allele to create a clinical-variant (CV) strain. Entire coding of dat-1 is replaced with SLC6A3 coding sequence containing an amino-acid change polymorphism that has been observed in patient populations. Upon request, a SLC6A3 line with client-specific polymorphism is made as a clinical variant allele.

 

RediMODEL knock-out in C. elegans | $845 each kit Buy Now

SLC6A3 - dopamine transporter

DISCRIPTION:

SLC6A3 is an integral membrane protein responsible for transporting the dopamine neurotransmitter from synaptic space back into presynaptic neurons. (Other names: DAT, DAT1, PKDYS).

DISEASES*:

Parkinsonism-Dystonia Infantile, Attention Deficit Hyperactivity Disorder, Schizophrenia, Parkinson Disease, Cocaine-Related Disorders, Anxiety Disorders, Hyperactive Behavior, Psychoses, Substance-Induced, Major Affective Disorder, Dementia, Nervous System Disorder, Paranoia, Amphetamine-Related Disorders, Tic Disorder, Memory Disorders, Micronuclei Chromosome-Defective, Tobacco Addiction Susceptibility, Nerve Degeneration.

* All disease associations scored above a 0.1 threshold from disgenet.com

Protein domain - slc6a3 dat-1.png

HOMOLOGY

Human Gene SLC6A3 Alignment         
C. elegans Gene dat-1 align
Identity 46%
Similarity 65%
rediMODEL-KO

KNOCK OUT of C. elegans gene

dat-1 is the homologous gene to SLC6A3. The dat-1 gene functions to control dopamine levels at the synapse. When dat-1 is expressed and active, it pumps dopamine from the extracellular space and into the cytosol. Deletion mutations in dat-1 cause a high rate of paralysis when exposed to the hypotonic solution. The canonical allele in dat-1 (ok157) contains a large deletion in the coding region and is thought to render a knock-out (KO) of gene function. The most easily measured phenotype for the KO is a Swimming Worm Induced Paralysis (SWIP) assay. To decrease the risk of background mutations, the KO strain is 4x outcrossed. The resulting outcrossed strain containing (ok157) allele is offered as the SLC6A3 RediMODEL-KO strain. The KO strain is confirmed by PCR and shipped in the RediMODEL kit format. Included in the kit is N2 wild-type strain as a reference control.


RediMODEL Kit Details:

Each kit arrives with 3 frozen tubes of SLC6A3 RediMODEL-KO and 3 tubes of N2 for control and all materials needed for 2+ days of growth.

PLEASE PUT FROZEN TUBES AT -80C UPON ARRIVAL.

Authentication:

Genotype and phenotype data provides a capacity for the researcher to monitor and retain authentication, as per NIH guidelines.

Genotype data:

PCR Assay

The RediMODEL-KO line is sequence verified and followed by PCR. The gel image in Genotype Data shows the expected PCR band pattern in an agarose gel for the primer set used to verify the RediMODEL kit.



amplicon size
wild type 500 bp
SLC6A3 RediMODEL-KI 900 bp
 

Phenotype data:

Bio Assay

The RediMODEL-KO line's activity is best followed by monitoring changes in thrashing behavior in hypotonic liquid. Mutants in thrashing behavior can be detected as number showing paralyssis upon timed exposure to liquid media. Using the Swimming Worm Induced Paralysis Protocol (P004), a significant difference in thrashing capacity is observed.

Swimming Worm Induced Paralysis Protocol (P004)

strain normal thrashing
N2 wild type 100%
SLC6A3 RediMODEL-KO 12%

Discover SLC6A3 RediMODEL-KO Kit: | $845 each kit Buy Now

Discover RediMODEL-KO Kit: | $845 each kit

Inquire on SLC6A3 RediMODEL-KO

Please provide contact info and a company representative will contact you soon with preorder invoicing details.

SLC6A3 rediMODEL-KO inquiry

RediMODEL knock-in of human gene replacement in C. elegans | $845 each kit Buy Now

SLC6A3 - dopamine transporter

DISCRIPTION:

SLC6A3 is an integral membrane protein responsible for transporting the dopamine neurotransmitter from synaptic space back into presynaptic neurons. (Other names: DAT, DAT1, PKDYS).

DISEASES*:

Parkinsonism-Dystonia Infantile, Attention Deficit Hyperactivity Disorder, Schizophrenia, Parkinson Disease, Cocaine-Related Disorders, Anxiety Disorders, Hyperactive Behavior, Psychoses, Substance-Induced, Major Affective Disorder, Dementia, Nervous System Disorder, Paranoia, Amphetamine-Related Disorders, Tic Disorder, Memory Disorders, Micronuclei Chromosome-Defective, Tobacco Addiction Susceptibility, Nerve Degeneration.

* All disease associations scored above a 0.1 threshold from disgenet.com

Protein domain - slc6a3 dat-1.png

HOMOLOGY

Human Gene SLC6A3 Alignment         
C. elegans Gene dat-1 align
Identity 46%
Similarity 65%
RediMODEL-KI logo sml.png

HUMANIZED C. elegans:

dat-1 is the homologous gene to SLC6A3. The dat-1 gene functions to control dopamine levels at the synapse. When dat-1 is expressed and active, it pumps dopamine from the extracellular space and into the cytosol. Deletion mutations in dat-1  as SLC6A3 RediMODEL-KO cause a high rate of paralysis when exposed to a hypotonic solution (SWIP - Swimming Worm Induced Paralysis) as a loss-of-function phenotype. To create a humanized worm, SLC6A3 gene is swapped for the dat-1 coding segment Installation of SLC6A3 in replacement of dat-1 causes restoration of SWIP phenotype and is offered as SLC6A3 RediMODEL-KI. The KI strain is confirmed by PCR and shipped in the RediMODEL kit format. Included in the kit is SLC6A3 RediMODEL-KO as a reference control.


RediMODEL Kit Details:

Each kit arrives with 3 frozen tubes of SLC6A3 RediMODEL-KI and 3 tubes of N2 for control and all materials needed for 2+ days of growth.

PLEASE PUT FROZEN TUBES AT -80C UPON ARRIVAL.

Authentication:

Genotype and phenotype data provides a capacity for the researcher to monitor and retain authentication, as per NIH guidelines.

Genotype data:

PCR Assay

The RediMODEL-KI line is sequence verified and followed by PCR. The gel image in Genotype Data shows the expected PCR band pattern in an agarose gel for the primer set used to verify the RediMODEL kit.



amplicon size
wild type 500 bp
SLC6A3 RediMODEL-KI 900 bp
 

Phenotype data:

Bio Assay

The RediMODEL-KI line's activity is best followed by monitoring changes in thrashing behavior in hypotonic liquid. Mutants in thrashing behavior can be detected as number showing paralyssis upon timed exposure to liquid media. Using the Swimming Worm Induced Paralysis Protocol (P004), a significant difference in thrashing capacity is observed.

Swimming Worm Induced Paralysis Protocol (P004)

strain normal thrashing
SLC6A3 RediMODEL-KO 12%
SLC6A3 RediMODEL-KI 100%

Discover SLC6A3 RediMODEL-KI Kit: | $845 each kit Buy Now

Discover RediMODEL-KI Kit: | $845 each kit

Inquire on SLC6A3 RediMODEL-KI

Please provide contact info and a company representative will contact you soon with preorder details.

SLC6A3 rediMODEL-KI inquiry

RediMODEL clinical variant in C. elegans | $6695 each kit Buy Now

SLC6A3 - dopamine transporter

DISCRIPTION:

SLC6A3 is an integral membrane protein responsible for transporting the dopamine neurotransmitter from synaptic space back into presynaptic neurons. (Other names: DAT, DAT1, PKDYS).

DISEASES*:

Parkinsonism-Dystonia Infantile, Attention Deficit Hyperactivity Disorder, Schizophrenia, Parkinson Disease, Cocaine-Related Disorders, Anxiety Disorders, Hyperactive Behavior, Psychoses, Substance-Induced, Major Affective Disorder, Dementia, Nervous System Disorder, Paranoia, Amphetamine-Related Disorders, Tic Disorder, Memory Disorders, Micronuclei Chromosome-Defective, Tobacco Addiction Susceptibility, Nerve Degeneration.

* All disease associations scored above a 0.1 threshold from disgenet.com

Protein domain - slc6a3 dat-1.png

HOMOLOGY

Human Gene SLC6A3 Alignment         
C. elegans Gene dat-1 align
Identity 46%
Similarity 65%
rediMODEL-KO

CLINICAL VARIANT in C. elegans:

dat-1 is the homologous gene to SLC6A3. The dat-1 gene functions to control dopamine levels at the synapse. When dat-1 is expressed and active, it pumps dopamine from the extracellular space and into the cytosol. Deletion mutation in dat-1  as the SLC6A3 RediMODEL-KO causes a high rate of paralysis when exposed to a hypotonic solution (SWIP - Swimming Worm Induced Paralysis) as a loss-of-function phenotype. Installation of SLC6A3 in replacement of dat-1 results in the SLC6A3 RediMODEL-KI wt-control strain and causes restoration of normal SWIP phenotype.  A client-specified clinical variant is installed into the SLC6A3 sequence of the wt-control strain.  The result is the SLC6A3 RediMODEL-CV strain whose relative phenotypic consequence can be rapidly measured. The CV strain is confirmed by PCR and shipped in the RediMODEL kit format. Included in the kit are SLC6A3 RediMODEL-KO and SLC6A3 RediMODEL-KI as reference controls.

genomic locus - SLC6A3 CV.png

RediMODEL Kit Details:

Each kit arrives with 3 frozen tubes of SLC6A3 RediMODEL-CV and 3 tubes of N2 for control and all materials needed for 2+ days of growth.

PLEASE PUT FROZEN TUBES AT -80C UPON ARRIVAL.

Authentication:

Genotype and phenotype data provides a capacity for the researcher to monitor and retain authentication, as per NIH guidelines.

Genotype data:

PCR Assay

The RediMODEL-CV line is sequence verified and followed by PCR. The gel image in Genotype Data shows the expected PCR band pattern in an agarose gel for the primer set used to verify the RediMODEL kit.



amplicon size
wild type 500 bp
SLC6A4 RediMODEL-CV 903 bp
 

Phenotype data:

Bio Assay

The RediMODEL-CV line's activity is best followed by monitoring changes in thrashing behavior in hypotonic liquid. Mutants in thrashing behavior can be detected as number showing paralyssis upon timed exposure to liquid media. Using the Swimming Worm Induced Paralysis Protocol (P004), a significant difference in thrashing capacity is observed.

Swimming Worm Induced Paralysis Protocol (P004)

strain normal thrashing
SLC6A3 RediMODEL-KO 12%
SLC6A3 RediMODEL-CV ??%
SLC6A3 RediMODEL-KI 100%

Discover SLC6A3 RediMODEL-CV Kit: | $6695 each kit Buy Now

Discover RediMODEL-CV Kit: | $6695 each kit

Inquire on SLC6A3 RediMODEL-CV

Please provide contact info and a company representative will contact you soon with design and invoicing details.

SLC6A3 rediMODEL-CV inquiry

Alignment of SLC6A3 to dat-1

Red asterisks indicate clinical variant alleles of pathogenic nature in humans that are conserved in C. elegans. As a result, 5 of the 9 known pathogenic alleles can be modeled in a rediMODEL-KI format. A CRISPR transgenesis approach can be used to make one of three mutations: G121S, V158I, L224P, G327R, T356M alleles.

Let us know if you want to model human disease in SLC6A3 with dat-1 mutations.