WORKFLOW KIT · CELL ENGINEERING

Engineered Cell Purification Kit

Co-deliver antibiotic resistance selection markers with your cargo to create a short selection window and enrich engineered cells.

Every kit includes: RNA · Optimized protocols · MicroBooster™ cartridges · Suggested readouts · Reagents

Upgrade Your Workflow

Enrich your engineered cell population by co-delivering a transient selection marker alongside your cargo of interest. After a brief selection step, cells that received the delivery are preferentially retained, resulting in a more purified population carrying your cargo while maintaining the original cell background.

The problem

Deliver a cargo to a pool of cells and only a fraction receive it; the rest dilute every downstream readout. The usual fix is to build a stable cell line with an integrated resistance cassette, which costs weeks of cloning and leaves a permanent selection marker in the genome. For a transient mRNA, protein, or RNP experiment, that is far more engineering than the question needs.

With this kit

Portal's technology enables co-delivery of antibiotic resistance markers with your cargo, this brief selection step purifies your population down to the cells that received the payload. Because the marker is mRNA, it fades on its own, leaving no integrated cassette or new cell line, just a purified version of the one you started with.

How the kits work

Gateway™ mechanoporation replaces the electroporation-plus-recovery step with a single, gentle delivery that the cells can be transplanted straight out of.

Portal has the Solution

01

Prepare

Your cells

02

Mix

mRNA + your cargo

03

Boost

Gateway™ mechanoporation

04

Select

Selection agent removes non-recipients

05

Enrich

Purified, cargo⁺ cells

Purified population, same cell line you started with

~16 hours

A brief puromycin selection clears unprotected cells, with no stable cell line needed

~10×

Fewer unprotected iPSCs recovered at 0.25 µg/mL puromycin, the lowest dose tested

90+%

Dextran-positive delivery in boosted iPSCs, so the marker co-delivers with your cargo

What's in the box

The cargo and protocols below make this kit specific. The core consumables, settings, and support ship with every Portal kit.

Available Cargoes

Puromycin-resistance (PuroR) mRNA
Custom RNA combinations
You provide: Your cargo of interest (mRNA, protein, RNP)

Don't see your cargo? Ask us about custom configurations.

Reagents & Consumables

Delivery tracer
MicroBooster™
Portal reagents

Protocol & Settings

Protocol for transient puromycin selection
Recommended cell inputs and puromycin dose
Suggested enrichment readouts

Support

Portal RNA sourcing
Optional: hands-on implementation support

Available Cargoes

Puromycin-resistance (PuroR) mRNA
Custom RNA combinations
You provide: Your cargo of interest (mRNA, protein, RNP)

Don't see your cargo? Ask us about custom configurations.

Reagents & Consumables

Delivery tracer
MicroBooster™
Portal reagents

Protocol & Settings

Protocol for transient puromycin selection
Recommended cell inputs and puromycin dose
Suggested enrichment readouts

Support

Portal RNA sourcing
Optional: hands-on implementation support

Proof Points

Enrichment of GFP+ Cells Following Transient Selection

iPSCs were co-delivered with GFP mRNA and a transient selection marker. One population underwent a brief purification step while the other remained unpurified. The purified population contained a higher proportion of GFP-positive cells.

Enrich Engineered Cells Through 3D Differentiation

iPSCs were engineered with GFP saRNA, NGN2 mRNA, and a transient selection marker, followed by embryoid body formation and selection during early 3D differentiation. The selection step preferentially removes non-engineered cells, enriching the remaining population for cargo-positive cells. Day 5 immunostaining shows GFP-positive cells throughout the aggregate alongside SOX2 and TUBB3 expression.

Check out our other kits

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Kits include: Optimized protocols · Portal reagents · Delivery cartridges · Suggested readouts

STEM CELL BIOLOGY

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Best suited for: Reprogramming, disease modeling

IMMUNOLOGY

APC Enhancement

Best suited for: PBMCs, dendritic cells
Available enhancements: mbIL2, mbIL12, CD86, custom antigen RNA
Best suited for: Immune activation, DC engineering, vaccine research

Cell Therapy

T Cell Functional Enhancement Kit

Best suited for: Human T cells (activated or unstimulated), PBMCs
Available enhancements: mb|L2, mbIL12, Perforin, Gzmb, B2M and TCR knockouts
Measure endogenous intracellular proteins in live cells
Use cases: Cytokine armoring, gene KO, persistence enhancement

NEUROSCIENCE

Neuronal Differentiation

Best suited for: iPSC-derived neurons, neural progenitors
Available enhancements: NGN2, ASCL1, LMX1A, FOXA2, NURR1, ASOs
Best suited for: Neuronal differentiation, disease modeling, Parkinson's/ALS

HSC Enhancement Kit

HSC Enhancement Kit

Best suited for: Hematopoietic stem cells
Available enhancements: CXCR4, BCL2
Best suited for: Engraftment, survival, stemness preservation

Broad Compatibility

Portal’s platform supports delivery to a diverse range of cell types and cargoes

Delivery materials

Validated cell types

mRNA, siRNA, saRNA

Proteins & Peptides

CRISPR RNPs

Small Molecules

Polymers

Nanoparticles

Antibodies

Virus

PBMCs

T cells

B cells

NK cells

iPSCs

Monocytes

RBCs

HSCs

Delivery materials

mRNA, siRNA, saRNA

Proteins & Peptides

CRISPR RNPs

Small Molecules

Polymers

Nanoparticles

Antibodies

Virus

Validated cell types

PBMCs

T cells

B cells

NK cells

iPSCs

Monocytes

RBCs

HSCs

Is the Engineered Cell Purification Kit for research or clinical use?The Engineered Cell Purification Kit is a Portal Bio workflow kit that co-delivers a puromycin-resistance (PuroR) mRNA together with your cargo in a single Gateway mechanoporation boost, then applies a brief puromycin selection to keep only the cells that received the delivery. Because the resistance marker is mRNA, it is transient and nothing integrates into the genome, so you enrich for delivered cells without building a stable cell line. You keep the cell line you started with, now enriched for the cells carrying your cargo.

How does transient puromycin selection with an mRNA marker work?Your cells are boosted with PuroR mRNA and your cargo of interest at the same time. Cells that received the boost express the PuroR mRNA and become resistant to puromycin for a short time. A brief puromycin selection then removes the cells that did not receive the marker and leaves the delivered cells behind. The mRNA is degraded over the following days, so the resistance fades on its own and no selection cassette is left in the genome.

Why use a transient mRNA marker instead of a stable resistance cassette?Building a stable cell line means integrating a resistance gene, cloning, and weeks of expansion, and it leaves a permanent selection marker in the genome. For a transient experiment, where an mRNA, protein, or RNP only needs to be present for a few days, that is far more engineering than the question needs. A co-delivered mRNA marker gives you the same enrichment in the pool of cells you already have, with no integration and no cell line to build.

How strong is the puromycin selection?In an iPSC kill-curve, the lowest concentration tested, 0.25 µg/mL, reduced the surviving cell count about tenfold versus no puromycin, from roughly 3.4 million to 0.3 million, and higher concentrations up to 2 µg/mL gave the same result. Cells without the resistance marker are cleared at a low concentration, so selection can stay brief; around 16 hours has been enough to clear unprotected iPSCs.

Does the resistance marker actually co-deliver with my cargo?Yes. PuroR mRNA has been carried into iPSCs in the same boost as a functional cargo, with over 90% of boosted cells delivery-positive by a co-delivered dextran tracer, against effectively none in the unboosted control, at about 65% viability. The cells that receive the tracer are the same cells that receive the resistance marker and your payload, so surviving selection tracks with having received the delivery.

What cargo can I pair with the PuroR marker?Anything Gateway delivers, including mRNA, self-amplifying RNA, protein, and ribonucleoprotein (RNP) complexes, on their own or in combination. The kit supplies the PuroR mRNA and can source custom RNA combinations; you provide your cargo of interest. Delivery is by mechanoporation rather than a chemical or viral carrier, so the marker and your cargo enter together in one boost.

What cell types does the kit work in?The selection data shown are in human iPSCs, and Gateway mechanoporation delivers into a broad range of cells, including primary cells and cell lines that are hard to transfect. Because sensitivity to puromycin varies by cell type, a Portal scientist tunes the puromycin concentration and the length of the selection for your cells.

Ready to run the Engineered Cell Purfication Kit?

Request the kit and a Portal scientist will spec it for your cells, cargo, and readout.