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Revision as of 04:33, 25 July 2008 (view source) Hmasood (Talk | contribs) ← Older edit		Revision as of 02:09, 30 October 2008 (view source) Katias (Talk | contribs) Newer edit →
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	<!--- The Mission, Experiments --->		<!--- The Mission, Experiments --->
	{{PrincetonHeader}}		{{PrincetonHeader}}
-	===Note===	+	== Parts Submitted to the Registry ==
-	If you choose to include a '''Parts Submitted to the Registry''' page, please list your parts here.  This is not necessary but it may be a nice list to keep track of.	+	==== pLV-TRE-Ngn1-linker-EYFP-2A-mKate-Ubc-Bla ====
		+
		+	* Description:A plasmid allowing rtTA-induced expression of the cell-fate regulator Ngn1 linked to an enhanced yellow fluorescent protein reporter, and a red fluorescent protein reporter tethered by a 2A linker. This construct allows for the verification of the 2A linker, with the ideal result of yellow-labeled Ngn1 remaining in the nucleus and red-labeled mKate being exported to the cytoplasm.
		+	* BioBrick Number:[http://partsregistry.org/wiki/index.php?title=Part:BBa_K076000 BBa_K076000]
		+
		+
		+	==== pLV-TRE-Sox17-Ubc-Bla ====
		+
		+	* Description: This is the backbone plasmid into which SfiI_Ngn1-linker-EYFP-2A-mKate_SfiI was inserted after digestion with SfiI.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076001 BBa_K076001]
		+
		+
		+	==== Ngn1 ====
		+
		+	* Description: Ngn1 is a transcription factor that promotes the differentiation of embryonic stem cells into neuronal precursors.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076002 BBa_K076002]
		+
		+
		+	==== Linker ====
		+
		+	* Description: This short sequence connects the Ngn1 and EYFP gene products, providing a translational fusion. This enables us to detect the location of Ngn1 by tracking the appearance of fluorescent yellow protein within the cell.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076003 BBa_K076003]
		+
		+
		+	==== EYFP ====
		+
		+	* Description: This is an enhanced yellow fluorescent reporter protein.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076004 BBa_K076004]
		+
		+
		+	==== 2A-linker ====
		+
		+	* Description: This is a protein linker sequence that allows one reporter protein, in our case EYFP, to be linked to another, in our case mKate, under certain conditions and cleaved under certain other conditions.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076005 BBa_K076005]
		+
		+
		+	==== mKate ====
		+
		+	* Description: This is a red fluorescent reporter protein, found to be more reliable than MCherry or DsRed2.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076006 BBa_K076006]
		+
		+
		+	==== pLV-TRE-Ngn1-linker-EYFP-Ubc-Bla ====
		+
		+	* Description: This plasmid only contains the fluorescently-labeled Ngn1, which enables the location of this neuronal transcription factor to be determined within the cell.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076007 BBa_K076007]
		+
		+
		+	==== D5R or DRD5 ====
		+
		+	* Description: This gene encodes a specific subunit of a dopamine receptor.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076008 BBa_K076008]
		+
		+
		+	==== Lbx1 ====
		+
		+	* Description: This gene codes for a GABAergic inhibitory neurotransmitter.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076009 BBa_K076009]
		+
		+
		+	==== GLRA1 ====
		+
		+	* Description: This gene codes for an inhibitory glycine receptor.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076010 BBa_K076010]
		+
		+
		+	==== T2A linker ====
		+
		+	* Description: This part is a protein linker allowing for translational fusion between two gene products.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076011 BBa_K076011]
		+
		+
		+	==== P2A linker ====
		+
		+	* Description: This part is a protein linker allowing for translational fusion between two gene products.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076012 BBa_K076012]
		+
		+
		+	==== pLV-TRE-Lbx1-T2A-GLRA1-P2A-D5R-Ubc-Bla  ====
		+
		+	* Description: This plasmid allows for the simultaneous production of Lbx1 [produces GABAergic neurons], GLRA1 [a glycine receptor], and D5R [a subunit of a particular dopamine receptor]. This makes the neurons able to respond to both a dopamine and a glycine input, and in turn produce a GABA output.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076013 BBa_K076013]
		+
		+
		+	==== Mash1 ====
		+
		+	* Description: This gene is a cell fate regulator that promotes the differentiation of embryonic stem cells into neuronal precursors.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076014 BBa_K076014]
		+
		+
		+	==== pLV-TRE-Mash1-IRES2-mKate  ====
		+
		+	* Description: This plasmid encourages differentiation of embryonic stem cells into neuronal precursors and marks the cells with the red-fluorescent mKate.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076015 BBa_K076015]
		+
		+
		+	==== Bleomycin ====
		+
		+	* Description: This gene is an antibiotic that allows for the selection of cells containing any construct that includes it.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076016 BBa_K076016]
		+
		+
		+	==== Ubc promoter ====
		+
		+	* Description: This is a stable constitutive promoter.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076017 BBa_K076017]
		+
		+
		+	==== pLV-TRE-D5R-Ubc-Bleo  ====
		+
		+	* Description: This plasmid allows bleomycin-selected cells to express the D5R dopamine receptor subunit upon induction with doxycycline.
		+	* BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076018 BBa_K076018]

---

Revision as of 02:09, 30 October 2008

Contents 1 Parts Submitted to the Registry 1.1 pLV-TRE-Ngn1-linker-EYFP-2A-mKate-Ubc-Bla 1.2 pLV-TRE-Sox17-Ubc-Bla 1.3 Ngn1 1.4 Linker 1.5 EYFP 1.6 2A-linker 1.7 mKate 1.8 pLV-TRE-Ngn1-linker-EYFP-Ubc-Bla 1.9 D5R or DRD5 1.10 Lbx1 1.11 GLRA1 1.12 T2A linker 1.13 P2A linker 1.14 pLV-TRE-Lbx1-T2A-GLRA1-P2A-D5R-Ubc-Bla 1.15 Mash1 1.16 pLV-TRE-Mash1-IRES2-mKate 1.17 Bleomycin 1.18 Ubc promoter 1.19 pLV-TRE-D5R-Ubc-Bleo

Parts Submitted to the Registry

pLV-TRE-Ngn1-linker-EYFP-2A-mKate-Ubc-Bla

• Description:A plasmid allowing rtTA-induced expression of the cell-fate regulator Ngn1 linked to an enhanced yellow fluorescent protein reporter, and a red fluorescent protein reporter tethered by a 2A linker. This construct allows for the verification of the 2A linker, with the ideal result of yellow-labeled Ngn1 remaining in the nucleus and red-labeled mKate being exported to the cytoplasm.
• BioBrick Number:[http://partsregistry.org/wiki/index.php?title=Part:BBa_K076000 BBa_K076000]

pLV-TRE-Sox17-Ubc-Bla

• Description: This is the backbone plasmid into which SfiI_Ngn1-linker-EYFP-2A-mKate_SfiI was inserted after digestion with SfiI.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076001 BBa_K076001]

Ngn1

• Description: Ngn1 is a transcription factor that promotes the differentiation of embryonic stem cells into neuronal precursors.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076002 BBa_K076002]

Linker

• Description: This short sequence connects the Ngn1 and EYFP gene products, providing a translational fusion. This enables us to detect the location of Ngn1 by tracking the appearance of fluorescent yellow protein within the cell.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076003 BBa_K076003]

EYFP

• Description: This is an enhanced yellow fluorescent reporter protein.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076004 BBa_K076004]

2A-linker

• Description: This is a protein linker sequence that allows one reporter protein, in our case EYFP, to be linked to another, in our case mKate, under certain conditions and cleaved under certain other conditions.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076005 BBa_K076005]

mKate

• Description: This is a red fluorescent reporter protein, found to be more reliable than MCherry or DsRed2.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076006 BBa_K076006]

pLV-TRE-Ngn1-linker-EYFP-Ubc-Bla

• Description: This plasmid only contains the fluorescently-labeled Ngn1, which enables the location of this neuronal transcription factor to be determined within the cell.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076007 BBa_K076007]

D5R or DRD5

• Description: This gene encodes a specific subunit of a dopamine receptor.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076008 BBa_K076008]

Lbx1

• Description: This gene codes for a GABAergic inhibitory neurotransmitter.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076009 BBa_K076009]

GLRA1

• Description: This gene codes for an inhibitory glycine receptor.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076010 BBa_K076010]

T2A linker

• Description: This part is a protein linker allowing for translational fusion between two gene products.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076011 BBa_K076011]

P2A linker

• Description: This part is a protein linker allowing for translational fusion between two gene products.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076012 BBa_K076012]

pLV-TRE-Lbx1-T2A-GLRA1-P2A-D5R-Ubc-Bla

• Description: This plasmid allows for the simultaneous production of Lbx1 [produces GABAergic neurons], GLRA1 [a glycine receptor], and D5R [a subunit of a particular dopamine receptor]. This makes the neurons able to respond to both a dopamine and a glycine input, and in turn produce a GABA output.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076013 BBa_K076013]

Mash1

• Description: This gene is a cell fate regulator that promotes the differentiation of embryonic stem cells into neuronal precursors.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076014 BBa_K076014]

pLV-TRE-Mash1-IRES2-mKate

• Description: This plasmid encourages differentiation of embryonic stem cells into neuronal precursors and marks the cells with the red-fluorescent mKate.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076015 BBa_K076015]

Bleomycin

• Description: This gene is an antibiotic that allows for the selection of cells containing any construct that includes it.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076016 BBa_K076016]

Ubc promoter

• Description: This is a stable constitutive promoter.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076017 BBa_K076017]

pLV-TRE-D5R-Ubc-Bleo

• Description: This plasmid allows bleomycin-selected cells to express the D5R dopamine receptor subunit upon induction with doxycycline.
• BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076018 BBa_K076018]

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