Difference between revisions of "WormBase Model:Construct"

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<pre>
 
<pre>
 
Drives_construct ?Construct XREF Driven_by_gene
 
Drives_construct ?Construct XREF Driven_by_gene
 
+
</pre>
  
 
==Transgene==
 
==Transgene==

Revision as of 00:38, 6 March 2014

back to WormBase_Models

Proposed model changes

Purpose: the technology of engineering mutations and gene replacement has been developed in C. elegans. With these new research tools, capture and display of the molecular information of these alleles needs to be updated. We propose a new class, Construct, to capture the specifics of the DNA tool used to perform the replacement or engineering, while the Variation model gets updated to record the engineering event itself and its impact on the genome. As a side benefit to the creation of the Construct model, we can use this new class to also capture the genomic arrays used create transgenes.

Variation

Proposed additions

?Variation
Variation_type 
Engineered_allele 
Variation_summary //to house final engineered construct
    Derived_from ?Construct XREF Engineered_variation
Identical_transgene Unique ?Transgene XREF Identical_variation
Method 
    HR //Homologous_recombination
    NHEJ //Non-homologous DNA end-joining, imprecise DNA repair
    MosSci
    TALENs
    CRISPR/Cas9
    ZFN-NHEJ repair //Zinc-finger nuclease
    ZFN-HR repair
Expr_pattern ?Expr_pattern XREF Variation #Evidence

notes on variation model changes

see discussion tab

Construct (new)

Class itself is new

?Construct // any single contiguous stretch of engineered DNA sequence
Public_name  ?Text
Other_name  ?Text
Summary   ?Text //genotype like [Pmyo-2::GFP]
Sequence_feature ?Feature XREF Construct
Driven_by ?Gene XREF Drives_transgene
Gene ?Gene XREF Transgene_product
Fusion_reporter ?Text //fluorescent proteins GFP, RFP, mCherry, etc.
Other_reporter ?Text //to add reporters, tags that aren’t included in model
Purification_tag ?Text //FLAG, HA, Myc, TAP, etc.  
Recombination_site ?Text //LoxP, FRT
Type_of_construct
    Chimera
    Domain_swap
    Engineered mutation
    Fusion
    Complex // complex changes (e.g. GFP fusion plus point mutations)  
    Transcriptional_fusion
    Translational_fusion 
    N-terminal_translational_fusion
    C-terminal_translational_fusion
    Internal_coding_fusion
Selection_marker     ?Text    //for elements stitched into contiguous sequence, coinjected elements will get their own construct ID, these will be joined together to create transgene 
Construction_summary  ?Text    //Backbone vector, mol bio 
     DNA_text ?text // for mapping to genome, can include entire construct sequence
Used_for
    Transgene_construction ?Transgene XREF Construct
    Engineered_variation ?Variation XREF Derived_from
    Topic_output_indicator ?WBProcess XREF marker_construct
    Expression_pattern ?Expr_pattern XREF Construct 
Reference ?Paper XREF Construct  
Person ?Person XREF Construct
Laboratory ?Laboratory #Lab_Location 
Remark ?Text #Evidence

notes on construct model

see discussion tab

Expression_pattern

proposed addition

Variation ?Variation XREF Expression_pattern
Construct ?Construct XREF Expression_pattern

==Gene==
proposed change
<pre>
Drives_Transgene ?Transgene XREF Driven_by_gene

changed to

Drives_construct ?Construct XREF Driven_by_gene

Transgene

Proposed changes: Many of the transgene tags have been moved to the proposed ?Construct model, the remaining tags as well as some additions are shown below

?Transgene      
Summary UNIQUE ?Text                               
Synonym ?Text       
Identical_variation Unique ?Variation XREF Identical_transgene //put in to unambiguously associate the allele/transgene object - see discussion
Construction //Strain_construction
     Construct       ?Construct XREF Transgene_construction
     Coinjection ?Construct XREF Transgene_construction
     Coinjection_other ?Text
     Integration_method UNIQUE ?Text     
     Integrated_from ?Transgene XREF Transgene_derivation                                               
     Laboratory ?Laboratory #Lab_Location    
     Author ?Author                   
Genetic_information                            
     Extrachromosomal                
     Integrated 
         Map ?Map  #Map_position  //needed for transgenes with no granular mapping, e.g., just mapped to a LG
Phenotype ?Phenotype XREF Transgene #Phenotype_info
Phenotype_not_observed ?Phenotype XREF Not_in_Transgene #Phenotype_info  
Used_for  
     Transgene_derivation ?Transgene XREF Integrated_from                                                   
     Expr_pattern ?Expr_pattern XREF Transgene  
     Marker_for   ?Text #Evidence 
     Gene_regulation ?Gene_regulation XREF Transgene 
     Interactor ?Interaction
     Topic_output_indicator ?WBProcess XREF marker_transgene
Associated_with                   
     Marked_rearrangement ?Rearrangement XREF By_transgene
     Strain ?Strain XREF Transgene 
Reference ?Paper XREF Transgene  
Species UNIQUE ?Species       
Remark ?Text #Evidence

Test data

LP132 nmy-2(cp7[nmy-2::gfp + LoxP unc-119(+) LoxP]) I; unc-119(ed3) I
Variation : "WBVar020000000"
Public_name "cp7"
Engineered_allele 
Variation_summary "[nmy-2::gfp + LoxP unc-119(+) LoxP]"
Derived_from "WBConstruct00000010"
Derived_from "WBConstruct00000011"
HR //homologous recombination

Construct : "WBConstruct00000010"
Summary "[nmy-2::gfp]
Driven_by "WBGene00003777"
Fusion_reporter "GFP"
N-terminal_translational_fusion

Construct : "WBConstruct00000011"
Summary "[LoxP unc-119(+) LoxP]
Recombination_site "LoxP"
Gene "WBGene00003777"



bus-50(e5000[T110E]) = An engineered missense mutation

bus-50(e5001[bus-50::gfp]) aka bus-50::gfp = An engineered fusion of GFP to the C-terminus of BUS-50 bus-50::gfp. 
Variation : "WBVar0200000001"
Public_name "e5001"
Engineered_allele 
Variation_summary "[bus-50::gfp]"
Derived_from "WBConstruct00000012"
CRISPR-Cas9

Construct : "WBConstruct00000012"
Summary "[bus-50::gfp]"
Gene "WBGene0020000001"
Fusion_reporter "GFP"
C-terminal_translational_fusion


bus-50(e5002[bus-50::gfp + loxP unc-119(+) loxP]) An engineered insertion of GFP plus the unc-119(+) selectable marker, flanked by loxP sites.
Variation : "WBVar020000003"
Public_name "e5002"
Engineered_allele 
Variation_summary "[bus-50::gfp + loxP unc-119(+) loxP]"
Derived_from "WBConstruct00000012"
Derived_from "WBConstruct00000011"
HR

Construct : "WBConstruct00000012"
Summary "[bus-50::gfp]"
Gene "WBGene0020000001"
Fusion_reporter "GFP"
C-terminal_translational_fusion


Construct : "WBConstruct00000011"
Summary "[LoxP unc-119(+) LoxP]
Recombination_site "LoxP"
Gene "WBGene00003777"



bus-50(e5003[bus-50::gfp +loxP]) aka bus-50(e5003) = bus-50(e5002) following Cre-mediated recombinase removal of unc-119(+) leaving a single loxP site
Variation : "WBVar020000004"
Public_name "e5003"
Engineered_allele 
Variation_summary "[bus-50::gfp + loxP]"
Derived_from "WBConstruct00000012"
Derived_from "WBConstruct00000011"
HR

Construct : "WBConstruct00000012"
Summary "[bus-50::gfp]"
Gene "WBGene0020000001"
Fusion_reporter "GFP"
C-terminal_translational_fusion

Construct : "WBConstruct00000011"
Summary "[LoxP unc-119(+) LoxP]
Recombination_site "LoxP"
Gene "WBGene00003777"


eIs2002 = eIs2002[unc-119::gfp] = eIs2002[unc-119::gfp, III:2992500]  Engineered insertions in apparent intergenic region with optional descriptors (nature of the insertion or position in the genome)
Variation : "WBVar020000005"
Public_name "eIs2002"
Engineered_allele 
Variation_summary "[unc-119::gfp]"
Derived_from "WBConstruct00000013"
Identical_transgene "WBTransgene00024514"
MosSci

Construct : "WBConstruct00000013"
Summary "[unc-119::gfp]"
Gene "WBGene0020000001"
Fusion_reporter "GFP"
Translational_fusion

Transgene : "WBTransgene00024514"
Public_name "eIs2002"
Summary "[unc-119::GFP]"
Construct "WBConstruct00000013"



ozIs909, or ozIs909[unc-119::mCherry *eIs2002] = Engineered changes to existing Is (or Si) insertions, which should receive new Is numbers using originating lab’s prefix. The original Is insertion is indicated in brackets with a preceding asterisk (*), in order to allow searches for all derivatives from a given insertion. In this case, an engineered change from GFP to mCherry in eIs2002
Variation : "WBVar020000006"
Public_name "ozIs909"
Engineered_allele 
Variation_summary "[unc-119::mCherry *eIs2002]"
Derived_from "WBConstruct00000015"
Identical_transgene "WBTransgene00024515"
MosSci

Construct : "WBConstruct00000015"
Summary "[unc-119::mCherry]"
Gene "WBGene0020000001"
Fusion_reporter "mCherry"
Translational_fusion

Transgene : "WBTransgene00024515"
Public_name "ozIs909"
Summary "[unc-119::mCherry]
Construct "WBConstruct00000015"

The following depicts how current transgenes would be redistributed into the proposed Construct and Transgene models

(Original)Transgene : "WBTransgene00000001"
Public_name	"adEx1256"
Summary	"[egl-19::sGFP-NLS + lin-15(+)]"
Reporter_product	"GFP"
Driven_by_gene	"WBGene00001187"
Strain	"DA1256"
Reference	"WBPaper00029359"
Reporter_type	"Transcriptional fusion"
Synonym	"[C48A7.1::gfp]"


(New)Transgene : "WBTransgene00000001"
Public_name	"adEx1256"
Summary	"[egl-19::sGFP-NLS + lin-15(+)]"
Construct "WBConstruct00000001"
Coinjection_other "lin-15(+)"
Extrachromosomal
Strain	"DA1256"
Reference	"WBPaper00029359"

(New)Construct : WBConstruct00000001
Public_name	"adEx1256"
Other_name	"[C48A7.1::gfp]"
Summary	"[egl-19::sGFP-NLS]"
Fusion_reporter	"GFP"
Driven_by_gene	"WBGene00001187"
Reference	"WBPaper00029359"
Transcriptional_fusion



(Original)Transgene : "WBTransgene00000011"
Public_name	"adIs1240"
Summary	"[lin-15(+) eat-4::sGFP]"
Reporter_product	"GFP"
Driven_by_gene	"WBGene00001135"
Strain	"DA1240"
Strain	"DA1243"
Map	"X"
Map_evidence	Paper_evidence	"WBPaper00038205"
Reference	"WBPaper00030960"
Reference	"WBPaper00032252"
Reference	"WBPaper00035265"
Reference	"WBPaper00036277"
Reference	"WBPaper00036704"
Reference	"WBPaper00037626"
Reference	"WBPaper00038205"
Reference	"WBPaper00044482"
Synonym	"[eat-4::gfp]"

(New)Transgene : "WBTransgene00000011"
Public_name	"adIs1240"
Summary	"[lin-15(+) eat-4::sGFP]"
Strain	"DA1240"
Strain	"DA1243"
Map	"X"
Map_evidence	Paper_evidence	"WBPaper00038205"
Coninjection_other "lin-15(+)"
Reference	"WBPaper00030960"
Reference	"WBPaper00032252"
Reference	"WBPaper00035265"
Reference	"WBPaper00036277"
Reference	"WBPaper00036704"
Reference	"WBPaper00037626"
Reference	"WBPaper00038205"
Reference	"WBPaper00044482"

(New)Construct : "WBConstruct00000011"
Public_name	"adIs1240"
Other_name	"[eat-4::gfp]"
Summary	"[eat-4::sGFP]"
Fusion_reporter	"GFP"
Driven_by_gene	"WBGene00001135"
Reference	"WBPaper00030960"


(Original) Transgene : "WBTransgene00000017"
Public_name	"ajIs1"
Summary	"[pgp-5::gfp]"
Coinjection_marker	"pRF4[rol-6(su1006)]"
Construction_summary	"Integrated from BC10030 sEx864."
Reporter_product	"GFP"
Driven_by_gene	"WBGene00003999"
Driven_by_gene	"WBGene00006767"
Integration_method	"Gamma_ray"
Integrated
Reference	"WBPaper00002968"
Reference	"WBPaper00031023"

(New) Transgene : "WBTransgene00000017"
Public_name	"ajIs1"
Integration_method	"Gamma_ray"
Integrated
Reference	"WBPaper00002968"
Reference	"WBPaper00031023"
Integrated_from "WBTransgene00002030"
Construction_summary	"Integrated from BC10030 sEx864."

(New) Transgene : "WBTransgene00002030"
Public_name "sEx864"
Synonym "[pgp-5::gfp]"
Synonym	"[C05A9.1::gfp]"
Summary "[rCesC05A9.1::GFP + pCeh361]"
Extrachromosomal
Construct "WBConstruct00000017"
Construct "WBConstruct00000018"
Construct "WBConstruct00000002"

(New) Construct : "WBConstruct00000018"
Public_name "pRF4"
Summary "[rol-6(su1006)]"
Gene "WBGene00004397"

(New) Construct : "WBConstruct00000017"
Public_name "[pgp-5::gfp]"
Summary	"[rCesC05A9.1::GFP]"
Reporter_product	"GFP"
Transcriptional_fusion
Driven_by_gene	"WBGene00003999"

(New) Construct : "WBConstruct00000002"
Public_name "pCeh361"
Summary "[pCeh::DPY-5]"
Construction_summary "A 3.3-kb Nco I fragment containing a predicted cuticle collagen gene was isolated from the cosmid F27C1, and cloned into the Nco I site of pGEM-5 to generate the clone pCeh361"
Reference	"WBPaper00027361"

(New) Transgene : "WBTransgene00000600"
Public_name	"hIs2"
Summary	"[DPY-5::GFP + rol-6(su1006) + pBluescript]
Construct	"WBConstruct00000003"//pCeh358
Coinjection	"WBConstruct00000004"//PCes1943
Coinjection	"WBConstruct00000005"//pBluescript KS
Construction_summary	"Transgenic animals were generated by microinjection of pCeh358 (5 ng/ul) and pBluescript KS (100 ng/ul), or in combination with 50 ng/ul pCes1943, which carries a dominant rol-6 mutation [rol-6(su1006)] used as a morphological marker for successful transformation"

(New) Construct : "WBConstruct00000003"
Public_name	"pCeh358"
Summary	"[dpy-5::gfp]"
Driven_by	"WBGene00001067"
Gene	"WBGene00001067"
Translational_fusion
Construction_summary	"The dpy-5::gfp reporter construct pCeh358 was generated by insertion of a 750 bp Sph I fragment from pCeh361 into the Sph I site of the gfp expression vector pPD95.69 (kindly provided by A. Fire). This fragment contains 5' sequences from an Sph I site in the polylinker of pCeh361 to a site 30 bp downstream from the predicted DPY-5 initiator methionine, resulting in an in-frame fusion of the first 12 codons of dpy-5 with gfp."
Reference	"WBPaper00027361"

(New) Construct : "WBConstruct00000004"
Public_name	"pCes1943"
Summary "[rol-6(su1006)]

(New) Construct : "WBConstruct00000005"
Public_name	"pBluescript KS"

Examples of two expression objects pertaining to sequence feature


Example 1:

Expr_pattern : "Expr11377"
Anatomy_term	"WBbt:0005813" Certain
Anatomy_term	"WBbt:0008588" Certain
Anatomy_term	"WBbt:0008589" Certain
Gene	"WBGene00001948"
Life_stage	"WBls:0000003"
Life_stage	"WBls:0000041"
Pattern	"enh-1 was preferentially active in the posterior C and D lineages in the embryo and in body wall musculature in the adult."
Reference	"WBPaper00032967"
Reporter_gene
Construct	"WBConstruct00000020"
Associated_feature "WBsf047531"

Construct : "WBConstruct00000020"
Public_name "hlh-1.enh-1::pPD107.94"
Summary	"[WBsf047531::pPD107.94]"
Sequence_feature "WBsf047531"
Fusion_reporter	"GFP"
Construction_summary "Enhancer region enh-1 for hlh-1 cloned into the (del)Pes-10 basal promoter vector pPD107.94"
Expression_pattern "Expr11377"
Reference	"WBPaper00032967"



Example 2:

Expr_pattern : "Expr11284"
Anatomy_term	"WBbt:0006894"
Gene	"WBGene00001185"
Pattern	"The distal enhancer activity was observed in P6.p and its descendants from the two-cell stage and increased with time. Distal enhancer activity persisted into much later stages than the proximal enhancer did."
Reference	"WBPaper00005841"
Reporter_gene
Construct	"WBConstruct00000021"
Associated_feature "WBsf919543"


Construct : "WBConstruct00000021"
Public_name "[egl-17.distal::pPD122.53]"
Summary	"[WBsf984567::pPD122.53]"
Sequence_feature "WBsf919543"
Fusion_reporter	"GFP"
Construction_summary "Distal egl-17 enhancer inserted into the pPD122.53 vector."
Expression_pattern "Expr11284"
Reference	"WBPaper00005841"



Expression objects described with reporter fusions that do not have a classical Ex transgene designation


Expr_pattern : "Expr11598"
Anatomy_term	"WBbt:0003681" Certain
Anatomy_term	"WBbt:0005772" Certain
Anatomy_term	"WBbt:0006749" Certain
Gene	"WBGene00001753"
Life_stage	"WBls:0000023"
Life_stage	"WBls:0000041"
Pattern	"gst-5 was expressed in intestine, pharynx, and circumpharyngeal neurons. Expression is seen from L1 to adult stages."
Reference	"WBPaper00037704"
Reporter_gene	"[GST-5::GFP] translational fusion."
Construct	"WBConstruct00000021"
Reporter_gene


Construct : "WBConstruct00000021"
Public_name	"GST-5::GFP"
Summary	"[GST-5::GFP]"
Driven_by	"WBGene00001753"
Fusion_reporter	"GFP"
Translational_fusion
Construction_summary "[GST-5::GFP] translational fusion."
Expression_pattern "Expr11598"
Reference	"WBPaper00037704"

Interaction/Regulation object contains sequence feature. I am not sure shall we add construct tag in our model? --Xiaodong


Interaction : "WBInteraction000502404"
Change_of_expression_level
Interactor_overlapping_gene      "WBGene00004077" Trans_regulator
Interactor_overlapping_gene      "WBGene00004077" Variation "WBVar00241166"
Interactor_overlapping_gene      "WBGene00009560" Trans_regulated
Interactor_overlapping_gene      "WBGene00009560" Expr_pattern "Expr4287"
Feature_interactor       "WBsf034247" Cis_regulator
Interaction_summary      "psa-3 expression was induced in T.p after the T cell division, and it accumulated in the posterior granddaughters. psa-3 expression in the T.p lineage was much lower in a pop-1 hypomorphic allele, q645."
Reporter_gene    "[psa-3::gfp], translational fusion"
Transgene       "WBTransgene00010102"
Transcriptional 
Positive_regulate        Anatomy_term "WBbt:0006996"
Positive_regulate        Anatomy_term "WBbt:0006997"
Positive_regulate        Anatomy_term "WBbt:0007000"
Positive_regulate        Anatomy_term "WBbt:0007007"
Positive_regulate        Anatomy_term "WBbt:0007012"
Positive_regulate        Anatomy_term "WBbt:0007016"
Paper    "WBPaper00027741"Remark   "POP-1 function is known to be required in T.p to determine the neural fate. POP-1 regulates psa-3 expression in T.p through the POP-1 binding site."