RediMODEL transgenics in C. elegans

SLC6A4 - serotonin transporter

GENE DESCRIPTION:

  • Function: SLC6A4 is an integral membrane protein responsible for transporting the serotonin neurotransmitter from synaptic space back into presynaptic neurons.
  • Disease: Genetic defects in SLC6A4 have been found to be highly associated with Depressive Disorder.
 

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

SLC6A4 RediMODEL KO: Generated in the C. elegans homolog mod-5 with deletion of coding sequence to create a knock-out (KO) strain. Loss of function from mod-5 deletion leads to increased neurotransmission due to serotonin build up at the synapse. Major phenotype readout is the observation of a 5-fold increase in serotonin dependent pumping of pharynx muscle. An all-coding deletion mutant is currently available as gene knock-out.

 

SLC6A4 RediMODEL KI: Generated in the C. elegans homolog mod-5 with human gene insertion to create a knock-in (KI) strain. Entire coding of mod-5 is replaced with wild-type SLC6A4 coding sequence. A SLC6A4 gene replacement is currently available as a humanized wt line.

 

SLC6A4 RediMODEL CV: Generated in the C. elegans homolog mod-5 with human gene insertion containing a clinical variant allele to create a clinical-variant (CV) strain. Entire coding of mod-5 is replaced with SLC6A4 coding sequence containing an amino-acid change polymorphism that has been observed in patient populations. Upon request, a SLC6A4 line with client-specific polymorphism is made as a clinical variant allele.

 

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

SLC6A4 - serotonin transporter

GENE DESCRIPTION:

SLC6A4 is an integral membrane protein responsible for transporting the serotonin neurotransmitter from synaptic space back into presynaptic neurons. (Other names: HTT, 5HTT, OCD1, SERT, 5-HTT, SERT1, hSERT, 5-HTTLPR).

GENETIC DISEASE:

Depressive disorder, Autistic Disorder, Anxiety Disorders, Major Depressive Disorder, Schizophrenia, Bipolar Disorder, Sudden infant death syndrome, Pulmonary Hypertension, Alcoholic Intoxication, Chronic Mood Disorders, Obsessive-Compulsive Disorder, Mental Depression, Personality Disorders, Substance-Related Disorders, Substance Withdrawal Syndrome, Amphetamine-Related Disorders, Memory Disorders, Cocaine-Related Disorders, Cognition Disorders, Asperger Syndrome.

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

HOMOLOGY

Human Gene SLC6A4 Alignment         
C. elegans Gene mod-5 align
Identity 46%
Similarity 65%
rediMODEL-KO

KNOCK OUT of C. elegans gene:

mod-5 is the homologous gene to SLC6A4. mod-5 functions to control serotonin levels at the synapse. When mod-5 is expressed and active, it pumps serotonin from the extracellular space into the cytosol. Mutations in mod-5 frequently are loss-of-function alleles. The canonical allele in mod-5 is (n3314), which contains a small deletion in the gene coding region and is thought to render a knock-out (KO) of gene function. The most easily measured phenotype for the KO is a high rate of pharynx pumping in absence of food or serotonin. Because the small deletion has the potential to retain function, a full deletion of the mod-5 locus is used instead.  To decrease the risk of background mutations, the KO strain is 4x outcrossed. The full deletion outcrossed allele is offered as the SLC6A4 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 SLC6A4 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 614 bp
SLC6A4 RediMODEL-KO 813 bp
 

Phenotype data:

Bio Assay

The RediMODEL-KO line's activity is best followed by monitoring changes in neurotransmission rates of food intake. Pharynx pumping increases and decreases are easily monitored in the ScreenChip system assay (Nemametrix.com). Using the No Stimulation Pharynx Pumping Protocol (P003), a significant difference in pumping rate is observed.

No Stimulation Pharynx Pumping Protocol (P003)

strain frequency (Hz) SEM
N2 wild type 0.095 +/- 0.035
SLC6A4 RediMODEL-KO 1.84 +/- 0.29

Discover SLC6A4 RediMODEL-KO Kit: | $845 each kit Buy Now

Discover RediMODEL-KO Kit: | $845 each kit

Inquire on SLC6A4 RediMODEL-KO

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SLC6A4 rediMODEL-KO inquiry

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

SLC6A4 - serotonin transporter

GENE DESCRIPTION:

SLC6A4 is an integral membrane protein responsible for transporting the serotonin neurotransmitter from synaptic space back into presynaptic neurons. (Other names: HTT, 5HTT, OCD1, SERT, 5-HTT, SERT1, hSERT, 5-HTTLPR).

GENETIC DISEASE:

Depressive disorder, Autistic Disorder, Anxiety Disorders, Major Depressive Disorder, Schizophrenia, Bipolar Disorder, Sudden infant death syndrome, Pulmonary Hypertension, Alcoholic Intoxication, Chronic Mood Disorders, Obsessive-Compulsive Disorder, Mental Depression, Personality Disorders, Substance-Related Disorders, Substance Withdrawal Syndrome, Amphetamine-Related Disorders, Memory Disorders, Cocaine-Related Disorders, Cognition Disorders, Asperger Syndrome.

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

Protein domain - slc6a3 dat-1.png

HOMOLOGY

Human Gene SLC6A4 Alignment         
C. elegans Gene mod-5 align
Identity 46%
Similarity 65%
RediMODEL-KI logo sml.png

HUMANIZED C. elegans:

mod-5 is the homologous gene to SLC6A4. The mod-5 gene functions to control serotonin levels at the synapse. When mod-5 is expressed and active, it pumps serotonin from the extracellular space into the cytosol. Deletion mutation in mod-5 as SLC6A4 RediMODEL-KO causes a pharynx pumping defect. To create a humanized worm,  SLC6A4 gene is swapped for the mod-5 coding segment.   Installation of SLC6A4 in replacement of mod-5 creates restoration of pharynx pumping defect phenotype and is offered as RediMODEL-KI .  The KI strain is confirmed by PCR and shipped in the RediMODEL kit format. Included in the kit is SLC6A4 RediMODEL-KO as a reference control.


RediMODEL Kit Details:

Each kit arrives with 3 frozen tubes of SLC6A4  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
SLC6A4 RediMODEL-KI 900 bp
 

Phenotype data:

Bio Assay

The RediMODEL-KI line's activity is best followed by monitoring changes in neurotransmission rates of food intake. Pharynx pumping increases and decreases are easily monitored in the ScreenChip system assay (Nemametrix.com). Using the Low Stimulation Pharynx Pumping Protocol (P001), a significant difference in pumping rate is observed.

Low Stimulation Pharynx Pumping Protocol (P001)

strain wt frequency (Hz)
SLC6A4 RediMODEL-KO 100%
SLC6A4 RediMODEL-KI 10%

Discover SLC6A4 RediMODEL-KI Kit: | $845 each kit Buy Now

Discover RediMODEL-KI Kit: | $845 each kit

Inquire on SLC6A4 RediMODEL-KI

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

SLC6A4 rediMODEL-KI inquiry

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

SLC6A4 - serotonin transporter

GENE DESCRIPTION:

SLC6A4 is an integral membrane protein responsible for transporting the serotonin neurotransmitter from synaptic space back into presynaptic neurons. (Other names: HTT, 5HTT, OCD1, SERT, 5-HTT, SERT1, hSERT, 5-HTTLPR).

GENETIC DISEASE:

Depressive disorder, Autistic Disorder, Anxiety Disorders, Major Depressive Disorder, Schizophrenia, Bipolar Disorder, Sudden infant death syndrome, Pulmonary Hypertension, Alcoholic Intoxication, Chronic Mood Disorders, Obsessive-Compulsive Disorder, Mental Depression, Personality Disorders, Substance-Related Disorders, Substance Withdrawal Syndrome, Amphetamine-Related Disorders, Memory Disorders, Cocaine-Related Disorders, Cognition Disorders, Asperger Syndrome.

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

Protein domain - slc6a3 dat-1.png

HOMOLOGY

Human Gene SLC6A4 Alignment         
C. elegans Gene mod-5 align
Identity 46%
Similarity 65%
rediMODEL-KO

CLINICAL VARIANT in C. elegans:

mod-5 is the homologous gene to SLC6A4. The mod-5 gene functions to control serotonin levels at the synapse. When mod-5 is expressed and active, it pumps serotonin from the extracellular space into the cytosol. Deletion mutation in mod-5 as SLC6A4 RediMODEL-KO causes a pharynx pumping defect.  Installation of SLC6A4 in replacement of mod-5 results in the SLC6A4 RediMODEL-KI wt-control strain and causes restoration of normal pharynx pumping phenotype.  Client-specified clinical variants are installed on the control strain to enable relative assessment of phenotypic consequence as the SLC6A4 RediMODEL-CV strain. A client-specified clinical variant is installed into the SLC6A4 sequence of the wt-control strain.  The result is the SLC6A4 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 SLC6A4 RediMODEL-KO and SLC6A4 RediMODEL-KI as reference controls.


RediMODEL Kit Details:

Each kit arrives with 3 frozen tubes of SLC6A4 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 900 bp
 

Phenotype data:

ScreenChip Assay

The RediMODEL-CV line's activity is best followed by monitoring changes in neurotransmission rates of the food intake. Pharynx pumping increases and decreases are easily monitored in ScreenChip system assay (Nemametrix.com). Using the Low Stimulation Pharynx Pumping Protocol (P001), a significant difference in pumping rate is observed.

Low Stimulation Pharynx Pumping Protocol (P001)

strain frequency (Hz)
SLC6A4 RediMODEL-KO 100%
SLC6A4 RediMODEL-CV ??%
SLC6A4 RediMODEL-KI 10%

Discover SLC6A4 RediMODEL-CV Kit: | $6695 each kit Buy Now

Discover RediMODEL-CV Kit: | $6695 each kit

Inquire on SLC6A4 RediMODEL-CV

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

SLC6A4 RediMODEL-CV inquiry

Alignment of SLC6A4 to mod-5

Red asterisks indicate clinical variant alleles of pathogenic nature in humans that are conserved in C. elegans. As a result, 3 of the 12 known pathogenic alleles can be modeled in a rediMODEL-KI format. A CRISPR transgenesis approach can be used to make one of three mutations: S293F, P339L, F465L alleles.

Let us know if you want to model human disease in SLC6A4 with mod-5 mutations.