cell reprogramming

Qiang

human stem cell potency ranges from [blank] to unipotent (progenitor)

totipotent

depending on donor age, the stem cells can be adult or [blank]

embryonic

limitations of cellular reprogramming include ethics issues and lack of sufficient [blank] of cells

amount

other limitations of cellular reprogramming are [blank] compatibility, isolation difficulties, and undefined stemness and differentiation potential

transplantation

Somatic Cell Nuclear Transfer-ESCs involves putting adult skin cells into unfertilized [blank]

oocyte

Somatic Cell Nuclear Transfer-ESCs form a blastocyst and stem cells can be taken from the [blank] cell layer

inner

dolly the sheep is an example of this cellular reprogramming strategy:

Somatic Cell Nuclear Transfer-ESCs

[blanks] involve reprogramming somatic cells into stem cells

human induced pluripotent stem cells (hiPSCs)

during cell-cell fusion BJ fibroblasts are combined with [blanks]

human embryonic (hES) cells

in cell-cell fusion BJ fibroblasts are transduced with [blank] while hES cells are transfected with GFP hygromycin

puromycin

[blank] creates a quadruploid but is only used as proof of concept

cell-cell fusion

Cell fate changed by enforced expression of [blank blanks] created iPSCs

transcription factors

in cell fate made iPSCs there are 4 key pluripotency genes: Oct4, Sox2, [blank], Klf4 which are known as yamanaka factors

c-Myc

while both the 1st and 2nd generation iPSCs use retrovirus only the 2nd generation had viable [blanks]

chimeras

2nd generation used a selection of [blank] cells instead of fbx15+ cells

nanog+

the 1st generation of iPSCs had [blank] errors while the 2nd did not

methylation

2nd generation allowed for turning on of [blank] genes specifically and not exogenous genes

endogenous

despite being efficient, [blanks] have the limitation that they can cause cancer due to genomic integration

retroviruses

the two major uses of iPSCs are disease modeling and [blank blank]

cell therapy

cell therapy can be used for personalized medicine or [blank] editing and correction

gene

[blank] gene editing involved 9BP cuts but is less specific and can introduce mutations

zinc finger nuclease (ZFN)

[blank] not does recognize nucleotides but proteins, is laborious

transcription activator-like effector nucleases (TALENs)

now 90% of gene editing is done by [blank] but is less specific than TALEN

CRISPR-Cas9

ZFN uses 3 ZF modules, 3BP each and x2 for specificity fused to a [blank]

nuclease

TALEN uses [x] modules, 1 BP each, and x2 for specificity fused to a nuclease

10+

CRISPR-cas9 involves one targeting [blank] bound by a nuclease

RNA

iPSC problems include use of viral vectors, low efficiency of reprogramming (0.02%-0.002%), and [blank] (20% of mice)

tumorigenesis

other iPSC problems are requiring efficient protocols, [blank] memory problems, and genomic instability

epigenetic

reprogrammed [blanks] are fast to make 2-4 weeks but only last for a relatively short time

neurons

iPSCs take 8-12 weeks to make but can be maintained for over [x] months

6

iPSCs have high tumorigenesis and [blank] efficiency while reprogrammed neurons have the reverse

low

the three major strategies are somatic cell nuclear transfer, stem cell - somatic cell fusion, and [blank]

enforced ectopic expression of transcription factors

cell replacement therapeutic strategies include iPSC differentiated cell transplantation, direct reprogramming based cell transplantation, and [blank]

in vivo direct reprogramming based on dispensable/adverse cell types