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	<!--- The Mission, Experiments --->		<!--- The Mission, Experiments --->
	{{PrincetonHeader}}		{{PrincetonHeader}}
		+	== Coding Parts Submitted to the Registry ==
		+	==== Ubc promoter ====
-	===Note===	+	* '''Description:''' This is a stable constitutive promoter.
		+	* '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076017 BBa_K076017]
-	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.	+
		+	==== Ngn1 ====
		+
		+	* '''Description:''' This 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]
		+
		+
		+	==== 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]
		+
		+
		+	==== 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]
		+
		+
		+	==== 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]
		+
		+
		+	==== T2A linker ====
		+
		+	* '''Description:''' This 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 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]
		+
		+
		+	==== 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]
		+
		+
		+	==== EYFP ====
		+
		+	* '''Description:''' This is an enhanced yellow fluorescent reporter protein.
		+	* '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076004 BBa_K076004]
		+
		+
		+	==== 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]
		+
		+
		+	==== 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]
		+
		+
		+	== Plasmids Submitted to the Registry ==
		+
		+
		+	==== 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]
		+
		+
		+	==== 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]
		+
		+
		+	==== pLV-TRE-Ngn1-linker-EYFP-2A-mKate-Ubc-Bla ====
		+
		+	* '''Description:''' This plasmid allows rtTA-induced expression of the cell-fate regulator Ngn1 linked to an enhanced yellow fluorescent protein (EYFP) reporter, and a red fluorescent protein (mKate) 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]
		+
		+
		+	==== 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]
		+
		+
		+	==== pLV-TRE-Lbx1-T2A-GLRA1-P2A-D5R-Ubc-Bla  ====
		+
		+	* '''Description:''' This plasmid allows for the simultaneous production of Lbx1 [produces GABAergic neurons], GLRA1 [produces a specific glycine receptor], and D5R [produced 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]

---

Latest revision as of 02:31, 30 October 2008

Contents 1 Coding Parts Submitted to the Registry 1.1 Ubc promoter 1.2 Ngn1 1.3 Mash1 1.4 Linker 1.5 2A linker 1.6 T2A linker 1.7 P2A linker 1.8 mKate 1.9 EYFP 1.10 D5R or DRD5 1.11 Lbx1 1.12 GLRA1 1.13 Bleomycin 2 Plasmids Submitted to the Registry 2.1 pLV-TRE-Mash1-IRES2-mKate 2.2 pLV-TRE-Ngn1-linker-EYFP-Ubc-Bla 2.3 pLV-TRE-Ngn1-linker-EYFP-2A-mKate-Ubc-Bla 2.4 pLV-TRE-Sox17-Ubc-Bla 2.5 pLV-TRE-D5R-Ubc-Bleo 2.6 pLV-TRE-Lbx1-T2A-GLRA1-P2A-D5R-Ubc-Bla

Coding Parts Submitted to the Registry

Ubc promoter

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

Ngn1

• Description: This 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]

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]

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]

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]

T2A linker

• Description: This 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 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]

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]

EYFP

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

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]

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]

Plasmids Submitted to the Registry

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]

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]

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

• Description: This plasmid allows rtTA-induced expression of the cell-fate regulator Ngn1 linked to an enhanced yellow fluorescent protein (EYFP) reporter, and a red fluorescent protein (mKate) 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]

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]

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

• Description: This plasmid allows for the simultaneous production of Lbx1 [produces GABAergic neurons], GLRA1 [produces a specific glycine receptor], and D5R [produced 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]

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