Base Cell - Nucleus

Overview¶

The Base Cell is Base Cytosol encapsulated in Base Membrane via Encapsulation: Phase Transfer. The Base Cell is deployed in a glucose outer solution and represents the default synthetic cell for the Nucleus distribution. Base Cells have red fluorescent membranes (Liss-Rhod PE) and express GFP over time if functioning correctly.

Reference Composition¶

Module Dependencies

Cytosol

Membrane

Outer Solution

The inner solution encapsulated into the Base Cell is Base Cytosol at reaction concentration, with pOpen-deGFP DNA added as a reporter.

Component	Input concentration	Final concentration	Volume for one reaction (µL)
SMix	3.33x	1x	12
PMix	15 mg/mL	1.80 mg/mL	4.8
Ribosomes	10 µM	1.8 µM	7.2
`pOpen-deGFP` DNA	124 nM	3 nM	0.95
tRNA	35 mg/mL	3.5 mg/ml	4
Optiprep	1.32 mg/µL	0.043 mg/µL	1.33
RNase Inhibitor	40 000 U/mL	2000 U/mL	2
Water			6.12
Total volume (µL)			40

Component	Target Percentage (%)	Molecular Weight (g/mol)	Stock concentration (mg/mL)	Volume to add (µL)
POPC	70	760.076	25	162.17
Cholesterol	29.95	386.654	50	17.65
Liss-Rhod PE	0.05	1301.71	1	4.96

See Base Membrane for the full membrane spec.

Component	Concentration
Glucose	850 mM

Expected Behavior¶

Base Cells express deGFP over time, starting dark and increasing in green fluorescence as translation proceeds. Kinetics data are reported in DevNote bnext-devnotes-base-cell-01.

Combined green (488 nm) and red (561 nm) fluorescence channels. First time point (labeled “t = 0”) corresponds to 30 min after preparation of the inner solution, reflecting the time required to prepare liposomes.

Process¶

Base Cells are assembled and encapsulated using Encapsulation: Phase Transfer.

Constituent Modules¶

Base Cytosol — inner solution, encapsulated at reaction concentration
Base Membrane — 70:30 POPC:cholesterol bilayer

Credits¶

Developed by Surendra Yadav (b.next).