Editing Protoplast (section)

==Uses for protoplasts==
[[File:Protoplast fusion.jpg|thumb|right|Fused protoplast (on left), containing both chloroplasts (from a leaf cell) as well as a  coloured vacuole (from a petal).]] 
Protoplasts can be used to study [[biological membrane|membrane]] biology, including the uptake of [[macromolecule]]s and  [[virus]]es . These are also used in [[somaclonal variation]].

Protoplasts are widely used for [[DNA]] [[Transformation (genetics)|transformation]] (for making [[genetically modified organism]]s), since the cell wall would otherwise block the passage of DNA into the cell.<ref name="Davey-2005">{{cite journal | vauthors = Davey MR, Anthony P, Power JB, Lowe KC | title = Plant protoplasts: status and biotechnological perspectives | journal = Biotechnology Advances | volume = 23 | issue = 2 | pages = 131–171 | date = March 2005 | pmid = 15694124 | doi = 10.1016/j.biotechadv.2004.09.008 }}</ref> In the case of plant cells, protoplasts may be regenerated into whole plants first by growing into a group of plant cells that develops into a [[Callus (cell biology)|callus]] and then by regeneration of shoots ([[caulogenesis]]) from the callus using [[plant tissue culture]] methods.<ref>{{cite journal | vauthors = Thorpe TA | title = History of plant tissue culture | journal = Molecular Biotechnology | volume = 37 | issue = 2 | pages = 169–180 | date = October 2007 | pmid = 17914178 | doi = 10.1007/s12033-007-0031-3 | s2cid = 25641573 }}</ref> Growth of protoplasts into callus and regeneration of shoots requires the proper balance of plant growth regulators in the tissue culture medium that must be customized for each species of plant.<ref>{{cite journal | vauthors = Sandgrind S, Li X, Ivarson E, Ahlman A, Zhu LH | title = Establishment of an Efficient Protoplast Regeneration and Transfection Protocol for Field Cress (<i>Lepidium campestre</i>) | journal = Frontiers in Genome Editing | volume = 3 | pages = 757540 | date = 16 November 2021 | pmid = 34870274 | pmc = 8635052 | doi = 10.3389/fgeed.2021.757540 | doi-access = free }}</ref><ref>{{cite journal | vauthors = Li X, Sandgrind S, Moss O, Guan R, Ivarson E, Wang ES, Kanagarajan S, Zhu LH | title = Efficient Protoplast Regeneration Protocol and CRISPR/Cas9-Mediated Editing of Glucosinolate Transporter (<i>GTR</i>) Genes in Rapeseed (<i>Brassica napus</i> L.) | journal = Frontiers in Plant Science | volume = 12 | pages = 680859 | date = 2021-07-07 | pmid = 34305978 | pmc = 8294089 | doi = 10.3389/fpls.2021.680859 | doi-access = free | bibcode = 2021FrPS...1280859L }}</ref> Unlike protoplasts from [[vascular plant]]s, protoplasts from [[moss]]es, such as ''[[Physcomitrella patens]]'', do not need [[phytohormone]]s for regeneration, nor do they form a callus during [[Regeneration (biology)|regeneration]]. Instead, they regenerate directly into the filamentous [[protonema]], mimicking a germinating moss spore.<ref>Bhatla SC, Kiessling J, [[Ralf Reski|Reski R]] (2002): [https://archive.today/20130203001100/http://www.springerlink.com/content/m747f7xfu9dxyprw/?p=2747894a7a9849a09e60b0e49f51afdc&pi=3 Observation of polarity induction by cytochemical localization of phenylalkylamine-binding receptors in regenerating protoplasts of the moss ''Physcomitrella patens'']. ''Protoplasma'' 219, 99–105.</ref>

Protoplasts may also be used for [[plant breeding]], using a technique called [[protoplast fusion]].  Protoplasts from different species are induced to fuse by using an [[electric field]] or a solution of [[polyethylene glycol]].<ref>{{cite journal | vauthors = Hain R, Stabel P, Czernilofsky AP, Steinbiß HH, Herrera-Estrella L, Schell J | title = Uptake, integration, expression and genetic transmission of a selectable chimaeric gene by plant protoplasts. | journal = Molecular and General Genetics MGG | date = May 1985 | volume = 199 | issue = 2 | pages = 161–168 | doi = 10.1007/BF00330254 }}</ref> This technique may be used to generate [[somatic hybrid]]s in tissue culture.{{Citation needed|date=August 2023}}

Additionally, protoplasts of plants expressing fluorescent proteins in certain cells may be used for Fluorescence Activated Cell Sorting (FACS), where only cells fluorescing a selected wavelength are retained. Among other things, this technique is used to isolate specific cell types (e.g., [[guard cells]] from leaves, [[pericycle]] cells from roots) for further investigations, such as transcriptomics.{{Citation needed|date=August 2023}}